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1. INFOGEST inter-laboratory recommendations for assaying gastric and pancreatic lipases activities prior to in vitro digestion studies

Author

Myriam M.L. Grundy, Evan Abrahamse, Annette Almgren, Marie Alminger, Ana Andres, Renata M.C. Ariëns, Shanna Bastiaan-Net, Claire Bourlieu-Lacanal, André Brodkorb, Maria R. Bronze, Irene Comi, Leslie Couëdelo, Didier Dupont, Annie Durand, Sedef N. El, Tara Grauwet, Christine Heerup, Ana Heredia, Marcos R. Infantes Garcia, Christian Jungnickel, Ilona E. Kłosowska-Chomiczewska, Marion Létisse, Adam Macierzanka, Alan R. Mackie, David J. McClements, Olivia Menard, Anne Meynier, Marie-Caroline Michalski, Ana-Isabel Mulet-Cabero, Anette Mullertz, Francina M. Payeras Perelló, Irene Peinado, Mélina Robert, Sébastien Secouard, Ana T. Serra, Sandra D. Silva, Gabriel Thomassen, Cecilia Tullberg, Ingrid Undeland, Carole Vaysse, Gerd E. Vegarud, Sarah H.E. Verkempinck, Michelle Viau, Mostafa Zahir, Ruojie Zhang, and Frédéric Carrière

Subjects
Lipases, Lipolysis, Enzyme activity, Titration method, Inhibitor, INFOGEST, Nutrition. Foods and food supply, TX341-641
Abstract

In vitro digestion studies often use animal digestive enzyme extracts as substitutes of human gastric and pancreatic secretions. Pancreatin from porcine origin is thus commonly used to provide relevant pancreatic enzymes such as proteases, amylase and lipase. Rabbit gastric extracts (RGE) have been recently introduced to provide gastric lipase in addition to pepsin. Before preparing simulated gastric and pancreatic extracts with targeted enzyme activities as described in in vitro digestion protocols, it is important to determine the activities of enzyme preparations using validated methods. The purpose of this inter-laboratory study within the INFOGEST network was to test the repeatability and reproducibility of lipase assays using the pH-stat technique for measuring the activities of gastric and pancreatic lipases from various sources. Twenty-one laboratories having different pH-stat devices received the same protocol with identical batches of RGE and two pancreatin sources. Lipase assays were performed using tributyrin as a substrate and three different amounts (50, 100 and 200 µg) of each enzyme preparation. The repeatability results within individual laboratories were satisfactory with coefficients of variation (CVs) ranging from 4 to 8% regardless of the enzyme amount tested. However, the inter-laboratory variability was high (CV > 15%) compared to existing standards for bioanalytical assays. We identified and weighted the contributions to inter-laboratory variability of several parameters associated with the various pH-stat equipment used in this study (e.g. reaction vessel volume and shape, stirring mode and rate, burette volume for the automated delivery of sodium hydroxide). Based on this, we established recommendations for improving the reproducibility of lipase assays using the pH-stat technique. Defining accurate and complete recommendations on how to correctly quantify activity levels of enzyme preparations is a gateway to promising comparison of in vitro data obtained from different laboratories following the same in vitro digestion protocol.

Published
2021
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2. A Trajectory of Discovery: Metabolic Regulation by the Conditionally Disordered Chloroplast Protein, CP12

Author

Cassy Gérard, Frédéric Carrière, Véronique Receveur-Bréchot, Hélène Launay, and Brigitte Gontero

Subjects
Calvin-Benson-Bassham cycle, conditionally disordered protein, history of modern science, metabolism regulation, moonlighting protein, protein-protein interaction, Microbiology, QR1-502
Abstract

The chloroplast protein CP12, which is widespread in photosynthetic organisms, belongs to the intrinsically disordered proteins family. This small protein (80 amino acid residues long) presents a bias in its composition; it is enriched in charged amino acids, has a small number of hydrophobic residues, and has a high proportion of disorder-promoting residues. More precisely, CP12 is a conditionally disordered proteins (CDP) dependent upon the redox state of its four cysteine residues. During the day, reducing conditions prevail in the chloroplast, and CP12 is fully disordered. Under oxidizing conditions (night), its cysteine residues form two disulfide bridges that confer some stability to some structural elements. Like many CDPs, CP12 plays key roles, and its redox-dependent conditional disorder is important for the main function of CP12: the dark/light regulation of the Calvin-Benson-Bassham (CBB) cycle responsible for CO2 assimilation. Oxidized CP12 binds to glyceraldehyde-3-phosphate dehydrogenase and phosphoribulokinase and thereby inhibits their activity. However, recent studies reveal that CP12 may have other functions beyond the CBB cycle regulation. In this review, we report the discovery of this protein, its features as a disordered protein, and the many functions this small protein can have.

Published
2022
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3. Postprandial bile acid levels in intestine and plasma reveal altered biliary circulation in chronic pancreatitis patients

Author

Lydie Humbert, Dominique Rainteau, Noshine Tuvignon, Claude Wolf, Philippe Seksik, René Laugier, and Frédéric Carrière

Subjects
cholangiohepatic shunt, exocrine pancreatic insufficiency, lipidomics, Biochemistry, QD415-436
Abstract

Bile acid (BA) secretion and circulation in chronic pancreatitis (CP) patients with exocrine pancreatic insufficiency (EPI) were investigated by simultaneously measuring postprandial levels of individual BAs in duodenal contents and blood plasma using LC-MS/MS. CP patients and healthy volunteers (HVs) were intubated with gastric and duodenal tubes prior to the administration of a test meal and continuous aspiration of duodenal contents. Pancreatic lipase outputs in CP patients were very low (0.7 ± 0.2 mg) versus HVs (116.7 ± 68.1 mg; P < 0.005), thus confirming the severity of EPI. Duodenal BA outputs were reduced in CP patients (1.00 ± 0.89 mmol; 0.47 ± 0.42 g) versus HVs (5.52 ± 4.53 mmol; 2.62 ± 2.14 g; P < 0.15). Primary to secondary BA ratio was considerably higher in CP patients (38.09 ± 48.1) than HVs (4.15 ± 2.37; P < 0.15), indicating an impaired transformation of BAs by gut microbiota. BA concentrations were found below the critical micellar concentration in CP patients, while a high BA concentration peak corresponding to gallbladder emptying was evidenced in HVs. Conversely, BA plasma concentration was increased in CP patients versus HVs suggesting a cholangiohepatic shunt of BA secretion. Alterations of BA circulation and levels may result from the main biliary duct stenosis observed in these CP patients and may aggravate the consequences of EPI on lipid malabsorption.

Published
2018
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4. Storage Compound Accumulation in Diatoms as Response to Elevated CO2 Concentration

Author

Erik L. Jensen, Karen Yangüez, Frédéric Carrière, and Brigitte Gontero

Subjects
co2, chrysolaminarin, diatoms, triacylglycerol, Biology (General), QH301-705.5
Abstract

Accumulation of reserve compounds (i.e., lipids and chrysolaminarin) in diatoms depends on the environmental conditions, and is often triggered by stress conditions, such as nutrient limitation. Manipulation of CO2 supply can also be used to improve both lipids and carbohydrates accumulation. Given the high diversity among diatoms, we studied the two marine model diatoms—Thalassiosira pseudonana and Phaeodactylum tricornutum, a freshwater diatom, Asterionella formosa, and Navicula pelliculosa—found in fresh- and sea-water environments. We measured the accumulation of reserve compounds and the activity of enzymes involved in carbon metabolism in these diatoms grown at high and atmospheric CO2. We observed that biomass and lipid accumulation in cells grown at high CO2 differ among the diatoms. Lipid accumulation increased only in P. tricornutum and N. pelliculosa grown in seawater in response to elevated CO2. Moreover, accumulation of lipids was also accompanied by an increased activity of the enzymes tested. However, lipid accumulation and enzyme activity decreased in N. pelliculosa cultured in fresh water. Chrysolaminarin accumulation was also affected by CO2 concentration; however, there was no clear relation with lipids accumulation. Our results are relevant to understand better the ecological role of the environment in the diatom adaptation to CO2 and the mechanisms underpinning the production of storage compounds considering diatom diversity.

Published
2019
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5. A broad pH range indicator-based spectrophotometric assay for true lipases using tributyrin and tricaprylin

Author

María de los Angeles Camacho-Ruiz, Juan Carlos Mateos-Díaz, Frédéric Carrière, and Jorge A. Rodriguez

Subjects
bile acids and salts, fatty acid, lipids, triglycerides, high-throughput screening, pH-dependent activity, Biochemistry, QD415-436
Abstract

A continuous assay is proposed for the screening of acidic, neutral, or alkaline lipases using microtiter plates, emulsified short- and medium-chain TGs, and a pH indicator. The lipase activity measurement is based on the decrease of the pH indicator optical density due to protonation which is caused by the release of FFAs during the hydrolysis of TGs and thus acidification. Purified lipases with distinct pH optima and an esterase were used to validate the method. The rate of lipolysis was found to be linear with time and proportional to the amount of enzyme added in each case. Specific activities measured with this microplate assay method were lower than those obtained by the pH-stat technique. Nevertheless, the pH-dependent profiles of enzymatic activity were similar with both assays. In addition, the substrate preference of each enzyme tested was not modified and this allowed discriminating lipase and esterase activities using tributyrin (low water solubility) and tricaprylin (not water soluble) as substrates. This continuous lipase assay is compatible with a high sample throughput and can be applied for the screening of lipases and lipase inhibitors from biological samples.

Published
2015
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6. Screening of phospholipase A activity and its production by new actinomycete strains cultivated by solid-state fermentation

Author

Priscila Sutto-Ortiz, María de los Angeles Camacho-Ruiz, Manuel R. Kirchmayr, Rosa María Camacho-Ruiz, Juan Carlos Mateos-Díaz, Alexandre Noiriel, Frédéric Carrière, Abdelkarim Abousalham, and Jorge A. Rodríguez

Subjects
Phospholipase A, Phospholipids, Actinomycetes, High-throughput screening, Solid-state fermentation, Medicine, Biology (General), QH301-705.5
Abstract

Novel microbial phospholipases A (PLAs) can be found in actinomycetes which have been poorly explored as producers of this activity. To investigate microbial PLA production, efficient methods are necessary such as high-throughput screening (HTS) assays for direct search of PLAs in microbial cultures and cultivation conditions to promote this activity. About 200 strains isolated with selected media for actinomycetes and mostly belonging to Streptomyces (73%) and Micromonospora (10%) genus were first screened on agar-plates containing the fluorophore rhodamine 6G and egg yolk phosphatidylcholine (PC) to detect strains producing phospholipase activity. Then, a colorimetric HTS assay for general PLA activity detection (cHTS-PLA) using enriched PC (≈60%) as substrate and cresol red as indicator was developed and applied; this cHTS-PLA assay was validated with known PLAs. For the first time, actinomycete strains were cultivated by solid-state fermentation (SSF) using PC as inductor and sugar-cane bagasse as support to produce high PLA activity (from 207 to 2,591 mU/g of support). Phospholipase activity of the enzymatic extracts from SSF was determined using the implemented cHTS-PLA assay and the PC hydrolysis products obtained, were analyzed by TLC showing the presence of lyso-PC. Three actinomycete strains of the Streptomyces genus that stood out for high accumulation of lyso-PC, were selected and analyzed with the specific substrate 1,2-α-eleostearoyl-sn-glycero-3-phosphocholine (EEPC) in order to confirm the presence of PLA activity in their enzymatic extracts. Overall, the results obtained pave the way toward the HTS of PLA activity in crude microbial enzymatic extracts at a larger scale. The cHTS-PLA assay developed here can be also proposed as a routine assay for PLA activity determination during enzyme purification,directed evolution or mutagenesis approaches. In addition, the production of PLA activity by actinomycetes using SSF allow find and produce novel PLAs with potential applications in biotechnology.

Published
2017
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7. Inhibition of CpLIP2 Lipase Hydrolytic Activity by Four Flavonols (Galangin, Kaempferol, Quercetin, Myricetin) Compared to Orlistat and Their Binding Mechanisms Studied by Quenching of Fluorescence

Author

Ruba Nasri, Luc P R Bidel, Nathalie Rugani, Véronique Perrier, Frédéric Carrière, Eric Dubreucq, and Christian Jay-Allemand

Subjects
recombinant CpLIP2 lipase, flavonols, orlistat, fluorescence quenching, inhibition, hydroxylation, tryptophan residues, docking, Organic chemistry, QD241-441
Abstract

The inhibition of recombinant CpLIP2 lipase/acyltransferase from Candida parapsiolosis was considered a key model for novel antifungal drug discovery and a potential therapeutic target for candidiasis. Lipases have identified recently as potent virulence factors in C. parapsilosis and some other yeasts. The inhibition effects of orlistat and four flavonols (galangin, kaempferol, quercetin and myricetin) characterized by an increasing degree of hydroxylation in B-ring, were investigated using ethyl oleate hydrolysis as the model reaction. Orlistat and kaempferol (14 µM) strongly inhibited CpLIP2 catalytic activity within 1 min of pre-incubation, by 90% and 80%, respectively. The relative potency of flavonols as inhibitors was: kaempferol > quercetin > myricetin > galangin. The results suggested that orlistat bound to the catalytic site while kaempferol interacted with W294 on the protein lid. A static mechanism of interactions between flavonols and CpLIP2 lipase was confirmed by fluorescence quenching analyses, indicating that the interactions were mainly driven by hydrophobic bonds and electrostatic forces. From the Lehrer equation, fractions of tryptophan accessibility to the quencher were evaluated, and a relationship with the calculated number of binding sites was suggested.

Published
2019
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8. An ultraviolet spectrophotometric assay for the screening of sn-2-specific lipases using 1,3-O-dioleoyl-2-O-α-eleostearoyl-sn-glycerol as substrate

Author

Lilia D. Mendoza, Jorge A. Rodriguez, Julien Leclaire, Gerard Buono, Frédéric Fotiadu, Frédéric Carrière, and Abdelkarim Abousalham

Subjects
Candida antarctica A lipase, β-cyclodextrin, stereoselectivity, sn-2 specificity, Aleurites fordii, tung oil, Biochemistry, QD415-436
Abstract

In the present study, we propose a continuous assay for the screening of sn-2 lipases by using triacylglycerols (TAGs) from Aleurites fordii seed (tung oil) and a synthetic TAG containing the α-eleostearic acid at the sn-2 position and the oleic acid (OA) at the sn-1 and sn-3 positions [1,3-O-dioleoyl-2-O-α-eleostearoyl-sn-glycerol (sn-OEO)]. Each TAG was coated into a microplate well, and the lipase activity was measured by optical density increase at 272 nm due to transition of α-eleostearic acid from the adsorbed to the soluble state. The sn-1,3-regioselective lipases human pancreatic lipase (HPL), LIP2 lipase from Yarrowia lipolytica (YLLIP2), and a known sn-2 lipase, Candida antarctica lipase A (CALA) were used to validate this method. TLC analysis of lipolysis products showed that the lipases tested were able to hydrolyze the sn-OEO and the tung oil TAGs, but only CALA hydrolyzed the sn-2 position. The ratio of initial velocities on sn-OEO and tung oil TAGs was used to estimate the sn-2 preference of lipases. CALA was the enzyme with the highest ratio (0.22 ± 0.015), whereas HPL and YLLIP2 showed much lower ratios (0.072 ± 0.026 and 0.038 ± 0.016, respectively). This continuous sn-2 lipase assay is compatible with a high sample throughput and thus can be applied to the screening of sn-2 lipases.

Published
2012
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9. Further biochemical characterization of human pancreatic lipase-related protein 2 expressed in yeast cells

Author

Cécilia Eydoux, Josiane De Caro, Francine Ferrato, Paul Boullanger, Dominique Lafont, René Laugier, Frédéric Carrière, and Alain De Caro

Subjects
constitutive expression, Pichia pastoris, inhibition, galactolipid, lid domain, galactolipase, Biochemistry, QD415-436
Abstract

Recombinant human pancreatic lipase-related protein 2 (rHPLRP2) was produced in the protease A-deficient yeast Pichia pastoris. A major protein with a molecular mass of 50 kDa was purified from the culture medium using SP-Sepharose and Mono Q chromatography. The protein was found to be highly sensitive to the proteolytic cleavage of a peptide bond in the lid domain. The proteolytic cleavage process occurring in the lid affected both the lipase and phospholipase activities of rHPLRP2. The substrate specificity of the nonproteolyzed rHPLRP2 was investigated using pH-stat and monomolecular film techniques and various substrates (glycerides, phospholipids, and galactolipids). All of the enzyme activities were maximum at alkaline pH values and decreased in the pH 5–7 range corresponding to the physiological conditions occurring in the duodenum. rHPLRP2 was found to act preferentially on substrates forming small aggregates in solution (monoglycerides, egg phosphatidylcholine, and galactolipids) rather than on emulsified substrates such as triolein and diolein. The activity of rHPLRP2 on monogalactosyldiglyceride and digalactosyldiglyceride monomolecular films was determined and compared with that of guinea pig pancreatic lipase-related protein 2, which shows a large deletion in the lid domain. The presence of a full-length lid domain in rHPLRP2 makes it possible for enzyme activity to occur at higher surface pressures. The finding that the inhibition of nonproteolyzed rHPLRP2 by tetrahydrolipstatin and diethyl-p-nitrophenyl phosphate does not involve any bile salt requirements suggests that the rHPLRP2 lid adopts an open conformation in aqueous media.

Published
2007
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10. A Metagenomic Investigation of the Duodenal Microbiota Reveals Links with Obesity.

Author

Emmanouil Angelakis, Fabrice Armougom, Frédéric Carrière, Dipankar Bachar, René Laugier, Jean-Christophe Lagier, Catherine Robert, Caroline Michelle, Bernard Henrissat, and Didier Raoult

Subjects
Medicine, Science
Abstract

Few studies have tested the small intestine microbiota in humans, where most nutrient digestion and absorption occur. Here, our objective was to examine the duodenal microbiota between obese and normal volunteers using metagenomic techniques.We tested duodenal samples from five obese and five normal volunteers using 16S rDNA V6 pyrosequencing and Illumina MiSeq deep sequencing. The predominant phyla of the duodenal microbiota were Firmicutes and Actinobacteria, whereas Bacteroidetes were absent. Obese individuals had a significant increase in anaerobic genera (p < 0.001) and a higher abundance of genes encoding Acyl-CoA dehydrogenase (p = 0.0018) compared to the control group. Obese individuals also had a reduced abundance of genes encoding sucrose phosphorylase (p = 0.015) and 1,4-alpha-glucan branching enzyme (p = 0.05). Normal weight people had significantly increased FabK (p = 0.027), and the glycerophospholipid metabolism pathway revealed the presence of phospholipase A1 only in the control group (p = 0.05).The duodenal microbiota of obese individuals exhibit alterations in the fatty acid and sucrose breakdown pathways, probably induced by diet imbalance.

Published
2015
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11. Continuous monitoring of cholesterol oleate hydrolysis by hormone-sensitive lipase and other cholesterol esterases

Author

Yassine Ben Ali, Frédéric Carrière, Robert Verger, Stefan Petry, Günter Muller, and Abdelkarim Abousalham

Subjects
cholesterol ester hydrolase, triacylglycerol lipase, cholesterol oleate emulsion, Biochemistry, QD415-436
Abstract

Hormone-sensitive lipase (HSL) contributes importantly to the hydrolysis of cholesteryl ester in steroidogenic tissues, releasing the cholesterol required for adrenal steroidogenesis. HSL has broad substrate specificity, because it hydrolyzes triacylglycerols (TAGs), diacylglycerols, monoacylglycerols, and cholesteryl esters. In this study, we developed a specific cholesterol esterase assay using cholesterol oleate (CO) dispersed in phosphatidylcholine and gum arabic by sonication. To continuously monitor the hydrolysis of CO by HSL, we used the pH-stat technique. For the sake of comparison, the hydrolysis of CO dispersion was also tested using other cholesteryl ester-hydrolyzing enzymes. The specific activities measured on CO were found to be 18, 100, 27, and 3 μmol/min/mg for HSL, cholesterol esterase from Pseudomonas species, Candida rugosa lipase-3, and cholesterol esterase from bovine pancreas, respectively. The activity of HSL on CO is ∼4- to 5-fold higher than on long-chain TAGs. In contrast, with all other enzymes tested, the rates of TAG hydrolysis were higher than those of CO hydrolysis. The relatively higher turnover of HSL on CO observed in vitro adds further molecular insight on the physiological importance of HSL in cholesteryl ester catabolism in vivo.Thus, HSL could be considered more as a cholesteryl ester hydrolase than as a TAG lipase.

Published
2005
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12. Sensitive assay for hormone-sensitive lipase using NBD-labeled monoacylglycerol to detect low activities in rat adipocytes

Author

Stefan Petry, Yassine Ben Ali, Henri Chahinian, Holger Jordan, Horst Kleine, Günter Müller, Frédéric Carrière, and Abdelkarim Abousalham

Subjects
p-nitrobenzofurazan, liposomes, NBD-fatty acid, Biochemistry, QD415-436
Abstract

The recent finding that p-nitrobenzofurazan (NBD)-FA is incorporated into and released from the acylglycerols of isolated rat adipocytes in an insulin-sensitive manner [G. Müller, H. Jordan, C. Jung, H. Kleine, and S. Petry. 2003. Biochimie. 85: 1245–1246] suggests that NBD-FA-labeled acylglycerols are cleaved by rat adipocyte hormone-sensitive lipase (HSL) in vivo. In the present study, we developed a continuous, sensitive in vitro lipase assay using a monoacylglycerol (MAG) containing NBD (NBD-MAG). NBD-MAG was found to provide an efficient substrate for rat adipocyte and human recombinant HSL. Ultrasonic treatment applied in the presence of phospholipids leads to the incorporation of NBD-MAG into the phospholipid liposomes and to a concomitant change of its spectrophotometric properties. The enzymatic release of NBD-FA and its dissociation from the carrier liposomes is accompanied by the recovery of the original spectrophotometric characteristics. The rate of lipolysis was monitored by measuring the increase in optical density at 481 nm, which was found to be linear with time and linearly proportional to the amount of lipase added. To assess the specific activity of recombinant HSL, we determined the molar extinction coefficient of NBD-FA under the assay conditions.This convenient assay procedure based on NBD-MAG should facilitate the search for small molecule HSL inhibitors.

Published
2005
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13. MmPPOX inhibits Mycobacterium tuberculosis lipolytic enzymes belonging to the hormone-sensitive lipase family and alters mycobacterial growth.

Author

Vincent Delorme, Sadia V Diomandé, Luc Dedieu, Jean-François Cavalier, Frédéric Carrière, Laurent Kremer, Julien Leclaire, Frédéric Fotiadu, and Stéphane Canaan

Subjects
Medicine, Science
Abstract

Lipid metabolism plays an important role during the lifetime of Mycobacterium tuberculosis, the causative agent of tuberculosis. Although M. tuberculosis possesses numerous lipolytic enzymes, very few have been characterized yet at a biochemical/pharmacological level. This study was devoted to the M. tuberculosis lipolytic enzymes belonging to the Hormone-Sensitive Lipase (HSL) family, which encompasses twelve serine hydrolases closely related to the human HSL. Among them, nine were expressed, purified and biochemically characterized using a broad range of substrates. In vitro enzymatic inhibition studies using the recombinant HSL proteins, combined with mass spectrometry analyses, revealed the potent inhibitory activity of an oxadiazolone compound, named MmPPOX. In addition, we provide evidence that MmPPOX alters mycobacterial growth. Overall, these findings suggest that the M. tuberculosis HSL family displays important metabolic functions, thus opening the way to further investigations linking the involvement of these enzymes in mycobacterial growth.

Published
2012
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14. The digestion of diacylglycerol isomers by gastric and pancreatic lipases and its impact on the metabolic pathways for TAG re-synthesis in enterocytes

Author

Jean-Claude Bakala-N’Goma, Leslie Couëdelo, Carole Vaysse, Marion Letisse, Véronique Pierre, Alain Géloen, Marie-Caroline Michalski, Michel Lagarde, Jean-David Leao, Frédéric Carrière, Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Equipe Nutrition, Santé et Biochimie des Lipides (ITERG), Cardiovasculaire, métabolisme, diabétologie et nutrition (CarMeN), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and ANR

Subjects
Absorption intestinale des graisses, Lipolyse, Enzyme, [SDV.IDA]Life Sciences [q-bio]/Food engineering, Lipides, Digestion, General Medicine, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Biochemistry
Abstract

International audience; Les activités spécifiques des lipases gastriques et pancréatiques ont été mesurées à l'aide de triacylglycérols (TAG) issus d'huile de colza, de 1,3-sn-DAG et de 1,2(2,3)-sn-DAG purifiés produits à partir de cette huile, ainsi que d'une huile de colza huile enrichie à 40% w/w DAG (DAGOIL). La lipase gastrique était plus active sur le 1,3 --DAG et le TAG, tandis que la lipase pancréatique présentait une sélectivité inverse avec une activité plus élevée sur le TAG que sur le DAG pris comme substrats initiaux. Cependant, dans les deux cas, les activités les plus élevées étaient affichées sur DAGOIL. Ces résultats montrent que le DAG mélangé au TAG, comme au cours de la digestion, est un meilleur substrat pour les lipases que le TAG. Les mêmes acylglycérols d'huile de colza ont été utilisés pour étudier l' absorption des graisses intestinales - sn -DAG que sur 1,2(2,3) -snchez le rat avec canulation du canal lymphatique mésentérique. Les taux de TAG synthétisés dans l'intestin et la concentration en acides gras totaux dans la lymphe n'étaient pas différents lorsque les rats recevaient des quantités identiques d'huile de colza TAG, de 1,2(2,3)-- DAG ou de DAGOIL. Depuis la lipolyse du 1,3- sn -DAG, 1,3- sn sn-DAG par les lipases digestives conduit au glycérol et non au 2-sn-monoacylglycérol (2- sn - MAG) comme la lipolyse des TAG, ces résultats suggèrent que la re-synthèse des TAG dans les entérocytes peut se faire entièrement via le « glycérol-3-phosphate ». (G3P) », avec la même efficacité que la voie 2- sn -MAG majoritairement impliquée dans l'absorption des graisses intestinales. Ces découvertes apportent un nouvel éclairage sur le rôle joué par les DAG en tant que produits intermédiaires de lipolyse. Selon leur structure, 1,2(2,3) -sn - DAG versus 1,3 - sn -DAG, les DAG peuvent contrôler la voie (2- sn -MAG ou G3P) par laquelle les TAG sont resynthétisés dans les entérocytes .

Published
2022
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15. Interfacial adsorption and activity of pancreatic lipase-related protein 2 onto heterogeneous plant lipid model membrane

Author

Jeanne Kergomard, Frédéric Carrière, Gilles Paboeuf, Lauriane Chonchon, Nathalie Barouh, Véronique Vié, Claire Bourlieu, Institut de Physique de Rennes (IPR), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro Montpellier, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Université de Montpellier (UM), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Démarche intégrée pour l'obtention d'aliments de qualité (UMR QualiSud), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Avignon Université (AU)-Université de La Réunion (UR)-Institut Agro Montpellier, Département Performances des systèmes de production et de transformation tropicaux (Cirad-PERSYST), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), and French Ministry of Research.

Subjects
liposomes, [PHYS]Physics [physics], heterogeneous monolayers, pancreatic lipase related-protein 2, galactolipids, monolayer, [SDV]Life Sciences [q-bio], General Medicine, [CHIM.MATE]Chemical Sciences/Material chemistry, Biochemistry, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition
Abstract

International audience; Pancreatic lipase related-protein 2 (PLRP2) exhibits remarkable galactolipase and phospholipase A1 activities, which depend greatly on the supramolecular organization of the substrates and the presence of surfactant molecules such as bile salts. The objective of the study was to understand the modulation of the adsorption mechanisms and enzymatic activity of Guinea pig PLRP2 (gPLRP2), by the physical environment of the enzyme and the physical state of its substrate. Langmuir monolayers were used to reproduce homogeneous and heterogeneous photosynthetic model membranes containing galactolipids (GL), and/or phospholipids (PL), and/or phytosterols (pS), presenting uncharged or charged interfaces. The same lipid mixtures were also used to form micrometric liposomes, and their gPLRP2 catalyzed digestion kinetics were investigated in presence or in absence of bile salts (NaTDC) during static in vitro, so called “bulk”, digestion.The enzymatic activity of gPLRP2 onto the galactolipid-based monolayers was characterized with an optimum activity at 15 mN/m, in the absence of bile salts. gPLRP2 showed enhanced adsorption onto biomimetic model monolayer containing negatively charged lipids. However, the compositional complexity in the heterogeneous uncharged model systems induced a lag phase before the initiation of lipolysis. In bulk, no enzymatic activity could be demonstrated on GL-based liposomes in the absence of bile salts, probably due to the high lateral pressure of the lipid bilayers. In the presence of NaTDC (4 mM), however, gPLRP2 showed both high galactolipase and moderate phospholipase A1 activities on liposomes, probably due to a decrease in packing and lateral pressure upon NaTDC adsorption, and subsequent disruption of liposomes.

Published
2023
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16. Digestibility and oxidative stability of plant lipid assemblies: An underexplored source of potentially bioactive surfactants?

Author

Claire Bourlieu, Véronique Vié, Nathalie Barouh, Jeanne Kergomard, Frédéric Carrière, Pierre Villeneuve, Institut de Physique de Rennes (IPR), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro - Montpellier SupAgro, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Démarche intégrée pour l'obtention d'aliments de qualité (UMR QualiSud), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Avignon Université (AU)-Université de La Réunion (UR)-Université de Montpellier (UM)-Institut Agro - Montpellier SupAgro, Département Performances des systèmes de production et de transformation tropicaux (Cirad-PERSYST), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)

Subjects
Metal chelating activity, Antioxidant, polar lipids, 030309 nutrition & dietetics, Digestibilité, medicine.medical_treatment, Composition des aliments, analyse des produits alimentaires, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Stabilité oxydative, surfactants, Industrial and Manufacturing Engineering, 03 medical and health sciences, Hydrolysis, Surfactant, medicine, [PHYS.COND]Physics [physics]/Condensed Matter [cond-mat], Q04 - Composition des produits alimentaires, Propriété physicochimique, digestive lipolysisantioxidants, 030304 developmental biology, [PHYS]Physics [physics], 2. Zero hunger, chemistry.chemical_classification, 0303 health sciences, Lipide, food and beverages, Galactolipids, Q01 - Sciences et technologies alimentaires - Considérations générales, General Medicine, bioactive properties [EN], Plant membrane, Chloroplast, Membrane, Biochemistry, chemistry, Composition (visual arts), [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Food Science, Polyunsaturated fatty acid
Abstract

International audience; Most lipids in our diet come under the form of triacylglycerols that are often redispersed and stabilized by surfactants in processed foods. In plant however, lipid assemblies constitute interesting sources of natural bioactive and functional ingredients. In most photosynthetic sources, polar lipids rich in ω3 fatty acids are concentrated. The objective of this review is to summarize all the knowledge about the physico-chemical composition, digestive behavior and oxidative stability of plant polar lipid assemblies to emphasize their potential as functional ingredients in human diet and their potentialities to substitute artificial surfactants/antioxidants. The specific composition of plant membrane assemblies is detailed, including plasma membranes, oil bodies, and chloroplast; emphasizing its concentration in phospholipids, galactolipids, peculiar proteins, and phenolic compounds. These molecular species are hydrolyzed by specific digestive enzymes in the human gastrointestinal tract and reduced the hydrolysis of triacylglycerols and their subsequent absorption. Galactolipids specifically can activate ileal break and intrinsically present an antioxidant (AO) activity and metal chelating activity. In addition, their natural association with phenolic compounds and their physical state (Lα state of digalactosyldiacylglycerols) in membrane assemblies can enhance their stability to oxidation. All these elements make plant membrane molecules and assemblies very promising components with a wide range of potential applications to vectorize ω3 polyunsaturated fatty acids, and equilibrate human diet.

Published
2021
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17. Interfacial adsorption of gastrointestinal lipases onto heterogenous biomimetic vegetal membranes

Author

Jeanne Kergomard, Frédéric Carrière, Gérard Lambeau, Gilles Paboeuf, Nathalie Barouh, Pierre Villeneuve, Claire Bourlieu, Véronique Vié, Institut de Physique de Rennes (IPR), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Institut de pharmacologie moléculaire et cellulaire (IPMC), Université Nice Sophia Antipolis (1965 - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), Démarche intégrée pour l'obtention d'aliments de qualité (UMR QualiSud), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Avignon Université (AU)-Université de La Réunion (UR)-Institut Agro Montpellier, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Université de Montpellier (UM), Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro Montpellier, and Kergomard, Jeanne

Subjects
[PHYS]Physics [physics], interfacial adsorption, [CHIM.ANAL] Chemical Sciences/Analytical chemistry, biomimetic vegetal lipid membrane, pancreatic sPLA2, lipolysis, [PHYS] Physics [physics], [SDV.AEN] Life Sciences [q-bio]/Food and Nutrition, gastric lipase, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, pancreatic lipase, [PHYS.COND]Physics [physics]/Condensed Matter [cond-mat], [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, [PHYS.COND] Physics [physics]/Condensed Matter [cond-mat]
Abstract

International audience; Langmuir films and their analysis by tensiometry, ellipsometry and Langmuir-Blodgett transfer coupled to atomic force microscopy are suitable tools to study the interactions of various lipases with model membranes. The objective of our study was to characterize the adsorption and lipolysis behavior of different gastrointestinal enzymes onto biomimetic vegetal membranes. An heterogenous monolayer based on a mixture of unsaturated galactolipids (MGDG, DGDG), saturated phospholipids (DPPC), and phytosterols was used. Four enzymes were studied, i) gastric lipase (GL), which has no lipolytic activity on polar lipids but whose adsorption may modulate the subsequent adsorption and activity of other lipases, and three pancreatic enzymes: ii) pancreatic lipase 2 (PLRP2) presenting both galactolipase and phospholipase A1 activities, (iii) pancreatic triacylglycerol lipase and its cofactor, colipase (PTL/coPTL), and (iv) pancreatic secreted phospholipase A2 (sPLA2-IB). A strong surfactant property of GL and its preferential adsorption at both the expanded lipid phase and at the boundary phase were observed, in line with previous results obtained with biomimetic milk fat globule membrane. With PLRP2, changes in surface pressure indicated a lipolytic activity in addition to lipase adsorption, with a higher rate at surface pressure below 15 mN/m. A drastic decrease in film thickness was also observed by ellipsometry, suggesting the removal of some lipolysis products. Such variation was not observed upon PTL/coPTL adsorption, and may be related to the absence of activity of PTL on polar lipids. Finally, the evolution of adsorption kinetic after injection of sPLA2-IB did not indicate a clear trend of lipolysis on the lipid film. However, a change in the morphology of condensed domains was observed by AFM, suggesting a phase separation of lipids induced by binding of sPLA2-IB.This study is a step forward in the understanding of interaction of gastrointestinal lipases with plant lipid membranes, an overlooked aspect of lipid digestion.

Published
2022

18. Bioaccessibility of essential lipophilic nutrients in a chloroplast-rich fraction (CRF) from agricultural green waste during simulated human gastrointestinal tract digestion

Author

Jutarat Wattanakul, Mansor Syamila, Randa Darwish, Mohamed A. Gedi, Poramat Sutcharit, Chao Chi, Patchaniya Akepach, Moulay Sahaka, Brigitte Gontero, Frédéric Carrière, David A. Gray, Bioénergétique et Ingénierie des Protéines (BIP ), and Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Chloroplasts, [SDV]Life Sciences [q-bio], Lutein, alpha-Tocopherol, food and beverages, Biological Availability, alpha-Linolenic Acid, General Medicine, Nutrients, beta Carotene, Gastrointestinal Tract, Humans, Digestion, Food Science
Abstract

The lipophilic nutrients in a chloroplast-rich fraction derived from pea vine postharvest field-residue are released in an in vitro digestion model; the extent of their release (bioaccessibility) is affected by heat-treatment of biomass or juice.

Published
2022
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19. Interfacial organization and phase behavior of mixed galactolipid-DPPC-phytosterol assemblies at the air-water interface and in hydrated mesophases

Author

Jeanne Kergomard, Frédéric Carrière, Gilles Paboeuf, Franck Artzner, Nathalie Barouh, Claire Bourlieu, Véronique Vié, Institut de Physique de Rennes (IPR), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro Montpellier, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Université de Montpellier (UM), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Démarche intégrée pour l'obtention d'aliments de qualité (UMR QualiSud), and Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Avignon Université (AU)-Université de La Réunion (UR)-Institut Agro Montpellier

Subjects
natural surfactant, F50 - Anatomie et morphologie des plantes, Colloid and Surface Chemistry, X-Ray Diffraction, Surfactant, Scattering, Small Angle, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Physical and Theoretical Chemistry, [PHYS.COND]Physics [physics]/Condensed Matter [cond-mat], Phytostérol, Propriété physicochimique, vegetal photosynthetic assemblies, phytosterol, [PHYS]Physics [physics], Lipide, Membrane cellulaire, Galactolipids, Membrane, Phytosterols, Water, Surfaces and Interfaces, General Medicine, Plants, Expérimentation en laboratoire, Plante, galactolipid, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Biotechnology
Abstract

International audience; The structural behavior of model assemblies composed of monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG), the two main galactolipids found in plants, was investigated at the air/water interface and in aqueous dispersion. To approach the composition of the natural photosynthetic membranes, tunable Langmuir model membrane of galactolipids (GL) were used, and were complexified to form either heterogenous binary or ternary assemblies of GL, phospholipids (PL), and phytosterols (pS).The impact of pS, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) or both on the structural properties of GL membrane was studied. The nature of the interactions between the different molecules was investigated using biophysical characterizations (ellipsometry, tensiometry, atomic force microscopy). In addition, the phase behavior was determined by SAXS analysis on the model assemblies in aqueous dispersions.Results revealed the good interfacial stability of these specific plant membrane lipids. The morphology of the GL film was characteristic of a fluid phase, with an interfacial roughness induced by the intercalation of monogalactosyl and digalactosyl polar heads of MGDG and DGDG, respectively. A phase heterogeneity in the monolayer was induced by the addition of DPPC and/or pS, which resulted in the modification of galactolipid organization and headgroup interactions. These structural changes were confirmed by SAXS analysis, showing more favorable interactions between MGDG and DPPC than between DGDG and DPPC in aqueous dispersion. This phenomenon was exacerbated in the presence of pS.HIGHLIGHTS • Plant polar lipids formed stable monolayers at the air/water interface. • Phytosterols reduced the lateral elasticity of galactolipid and DPPC monolayer. • DPPC has affected MGDG-DGDG interactions. • Results highlighted preferential phase miscibility between DPPC and MGDG.

Published
2022
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20. Cleaner degreasing of sheepskins by the Yarrowia lipolytica LIP2 lipase as a chemical-free alternative in leather industry

Author

Emna Moujehed, Zied Zarai, Haifa Khemir, Neila Miled, Mohamed Salah Bchir, Corinne Gablin, François Bessueille, Anne Bonhommé, Didier Leonard, Frédéric Carrière, Ahmed Aloulou, LGBEL - Laboratory of Biochemistry and Enzymatic Engineering of Lipases, Université de Sfax, Surfaces, Institut des Sciences Analytiques (ISA), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Natl Ctr Leather & Footwear, SO SA CUIR Tanning Co, Higher Inst Biotechnol Sfax, Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Emna Moujehed's doctoral work is supported by the National Program of Research and Innovation (PNRI) & the Ministry of Higher Education and Scientific Research, Tunisia., University of Sfax, National Engineering School of Sfax, Institut de Chimie du CNRS (INC)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)

Subjects
0303 health sciences, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, 05 social sciences, Leather industry, Yarrowia, Lipase, Surfaces and Interfaces, General Medicine, [CHIM.MATE]Chemical Sciences/Material chemistry, Fungal Proteins, 03 medical and health sciences, Colloid and Surface Chemistry, Yarrowia lipolytica LIP2 lipase, Enzymatic degreasing, 050501 criminology, [CHIM]Chemical Sciences, Physical and Theoretical Chemistry, 030304 developmental biology, 0505 law, Biotechnology
Abstract

Conventional degreasing of skins and hides in the leather industry requires high amounts of organic solvents and detergents that cause environmental issues. In this study, the LIP2 lipase from the yeast Yarrowia lipolytica (YLLIP2) was shown to be effective in degreasing sheepskins, thus reducing the amount of harmful chemicals. Using 6 mg of lipase/kg of raw skin, successful degreasing was achieved in only 15 min at pH 8 and 30°C. ToF-SIMS mass spectra of chemically and enzymatically treated sheepskins are consistent with a selective elimination process for the enzymatic treatment. Comparative SEM microscopy, ATR-FTIR spectroscopy and physicochemical analyses showed better properties of the enzymatically treated leather than those of the chemical treatment. Effluent physicochemical parameters showed that the enzymatic treatment is a cleaner degreasing operation. Altogether, this work opens new horizons to use the YLLIP2 lipase as a more efficient alternative in the leather industry.

Published
2022
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21. Targeting TOR signaling for enhanced lipid productivity in algae

Author

Marie-Hélène Montané, Laura Prioretti, Frédéric Carrière, Ben Field, Luisana Avilan, Brigitte Gontero, Benoît Menand, Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Luminy Génétique et Biophysique des Plantes (LGBP), Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) (BIAM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), ANR-15-CE05-0021,SIGNAUXBIONRJ,Manipulation des voies de signalisation de l'énergie afin d'améliorer la production de .lipides chez les eucaryotes photosynthétiques(2015), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, [SDV]Life Sciences [q-bio], Morpholines, Biochemistry, 03 medical and health sciences, Basic knowledge, Algae, Microalgae, Homeostasis, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Phaeodactylum tricornutum, Protein Kinase Inhibitors, Cholesterol homeostasis, Triglycerides, 030102 biochemistry & molecular biology, biology, TOR Serine-Threonine Kinases, Algal Proteins, General Medicine, Lipid Metabolism, biology.organism_classification, diatom, TOR signaling, Cell biology, 030104 developmental biology, Gene Expression Regulation, Productivity (ecology), Kinase inhibitors, Adipogenesis, Target of rapamycin, Biofuels, Lipogenesis, Signal Transduction
Abstract

International audience; Microalgae can produce large quantities of triacylglycerols (TAGs) and other neutral lipids that are suitable for making biofuels and as feedstocks for green chemistry. However, TAGs accumulate under stress conditions that also stop growth, leading to a trade-off between biomass production and TAG yield. Recently, in the model marine diatom Phaeodactylum tricornutum it was shown that inhibition of the target of rapamycin (TOR) kinase boosts lipid productivity by promoting TAG production without stopping growth. We believe that basic knowledge in this emerging field is required to develop innovative strategies to improve neutral lipid accumulation in oleaginous microalgae. In this minireview, we discuss current research on the TOR signaling pathway with a focus on its control on lipid homeostasis. We first provide an overview of the well characterized roles of TOR in mammalian lipogenesis, adipogenesis and lipolysis. We then present evidence of a role for TOR in controlling TAG accumulation in microalgae, and draw parallels between the situation in animals, plants and microalgae to propose a model of TOR signaling for TAG accumulation in microalgae.

Published
2020
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22. The digestion of galactolipids and its ubiquitous function in Nature for the uptake of the essential α-linolenic acid

Author

Noelia Aldai, Jutarat Wattanakul, Sawsan Amara, David Gray, Brigitte Gontero, Frédéric Carrière, Moulay Sahaka, John T. Christeller, Jérôme Lecomte, Pierre Villeneuve, Mohamed A. Gedi, Goetz Parsiegla, Bioénergétique et Ingénierie des Protéines (BIP ), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Lipolytech, University of Nottingham, School of Biosciences, University of Nottingham, UK (UON), University of the Basque Country, Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Département Performances des systèmes de production et de transformation tropicaux (Cirad-PERSYST), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Plant & Food Research, Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro - Montpellier SupAgro, and Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)

Subjects
0106 biological sciences, 0301 basic medicine, Insecta, Fonction physiologique, Protein Conformation, [SDV]Life Sciences [q-bio], 01 natural sciences, Carboxylesterase, MGDG, Galactolipase, Vegetables, Galactolipase activity, chemistry.chemical_classification, Chemistry, Hydrolysis, Fatty Acids, Fishes, alpha-Linolenic Acid, Galactolipids, General Medicine, Lipids, Milk, Biochemistry, Digestion, Lipid digestion, DGDG, Polyunsaturated fatty acid, Meat, Galactolipid, literature [EN], 03 medical and health sciences, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, Herbivory, Horses, Pancreas, L02 - Alimentation animale, Galactose, Fatty acid, Glycolipide, Lipase, Gastrointestinal Tract, Plant Leaves, 030104 developmental biology, Acide linolénique, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Carboxylic Ester Hydrolases, PUFA, 010606 plant biology & botany, Food Science
Abstract

International audience; Galactolipids, mainly monogalactosyl diglycerides and digalactosyl diglycerides are the main lipids found in the membranes of plants, algae and photosynthetic microorganisms like microalgae and cyanobacteria. As such, they are the main lipids present at the surface of the earth. They may represent up to 80 % of the fatty acid stocks, including a large proportion of polyunsaturated fatty acids mainly -linolenic acid (ALA). Nevertheless, the interest in these lipids for nutrition and other applications remains overlooked, probably because they are dispersed in the biomass and are not as easy to extract as vegetable oils from oleaginous fruit and oil seeds. Another reason is that galactolipids only represent a small fraction of the acylglycerolipids present in modern human diet. In herbivores such as horses, fish and folivorous insects, galactolipids may however represent the main source of dietary fatty acids due to their dietary habits and digestion physiology. The development of galactolipase assays has led to the identification and characterization of the enzymes involved in the digestion of galactolipids in the gastrointestinal tract, as well as by microorganisms. Pancreatic lipase-related protein 2 (PLRP2) has been identified as an important factor of galactolipid digestion in humans, together with pancreatic carboxyl ester hydrolase (CEH). The levels of PLRP2 are particularly high in monogastric herbivores thus highlighting the peculiar role of PLRP2 in the digestion of plant lipids. Similarly, pancreatic lipase homologs are found to be expressed in the midgut of folivorous insects, in which a high galactolipase activity can be measured. In fish, however, CEH is the main galactolipase involved. This review discusses the origins and fatty acid composition of galactolipids and the physiological contribution of galactolipid digestion in various species. This overlooked aspect of lipid digestion ensures not only the intake of ALA from its main natural source, but also the main lipid source of energy for growth of some herbivorous species.

Published
2020
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23. Reduction in Phosphoribulokinase Amount and Re-Routing Metabolism in Chlamydomonas reinhardtii CP12 Mutants

Author

Launay, Cassy Gérard, Régine Lebrun, Erwan Lemesle, Luisana Avilan, Kwang Suk Chang, EonSeon Jin, Frédéric Carrière, Brigitte Gontero, and Hélène

Subjects
Calvin–Benson–Bassham, CP12, enzymatic activity, intrinsically disordered protein, quantitative proteomics, photosynthesis, glyoxylate pathway, carbon allocation, nitrogen uptake
Abstract

The chloroplast protein CP12 is involved in the dark/light regulation of the Calvin–Benson–Bassham cycle, in particular, in the dark inhibition of two enzymes: glyceraldehyde−3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK), but other functions related to stress have been proposed. We knocked out the unique CP12 gene to prevent its expression in Chlamydomonas reinhardtii (ΔCP12). The growth rates of both wild-type and ΔCP12 cells were nearly identical, as was the GAPDH protein abundance and activity in both cell lines. On the contrary, the abundance of PRK and its specific activity were significantly reduced in ΔCP12, as revealed by relative quantitative proteomics. Isolated PRK lost irreversibly its activity over-time in vitro, which was prevented in the presence of recombinant CP12 in a redox-independent manner. We have identified amino acid residues in the CP12 protein that are required for this new function preserving PRK activity. Numerous proteins involved in redox homeostasis and stress responses were more abundant and the expressions of various metabolic pathways were also increased or decreased in the absence of CP12. These results highlight CP12 as a moonlighting protein with additional functions beyond its well-known regulatory role in carbon metabolism.

Published
2022
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24. Reduction in Phosphoribulokinase Amount and Re-Routing Metabolism in

Author

Cassy, Gérard, Régine, Lebrun, Erwan, Lemesle, Luisana, Avilan, Kwang Suk, Chang, EonSeon, Jin, Frédéric, Carrière, Brigitte, Gontero, and Hélène, Launay

Subjects
Phosphotransferases (Alcohol Group Acceptor), Glyceraldehyde-3-Phosphate Dehydrogenases, Photosynthesis, Chlamydomonas reinhardtii
Abstract

The chloroplast protein CP12 is involved in the dark/light regulation of the Calvin-Benson-Bassham cycle, in particular, in the dark inhibition of two enzymes: glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK), but other functions related to stress have been proposed. We knocked out the unique

Published
2022

25. Modulation of gastric lipase adsorption onto mixed galactolipid-DPPC films by the addition of phytosterols

Author

Jeanne Kergomard, Frédéric Carrière, Paboeuf, G., Franck Artzner, Nathalie Barouh, Pierre Villeneuve, Claire Bourlieu, Vié, V., Institut de Physique de Rennes (IPR), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS), Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Bioénergétique et Ingénierie des Protéines (BIP ), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Démarche intégrée pour l'obtention d'aliments de qualité (UMR QualiSud), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Avignon Université (AU)-Université de La Réunion (UR)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), Département Performances des systèmes de production et de transformation tropicaux (Cirad-PERSYST), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Kergomard, Jeanne, Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro - Montpellier SupAgro, Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), and Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Avignon Université (AU)-Université de La Réunion (UR)-Université de Montpellier (UM)-Institut Agro - Montpellier SupAgro

Subjects
[PHYS]Physics [physics], [SDV.AEN] Life Sciences [q-bio]/Food and Nutrition, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [PHYS.COND]Physics [physics]/Condensed Matter [cond-mat], [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, [PHYS.COND] Physics [physics]/Condensed Matter [cond-mat], ComputingMilieux_MISCELLANEOUS, [PHYS] Physics [physics]
Abstract

International audience

Published
2021

26. Fatty Acid Photodecarboxylase Is an Interfacial Enzyme That Binds to Lipid-Water Interfaces to Access Its Insoluble Substrate

Author

Cyril Aselmeyer, Frédéric Carrière, Anaïs Bénarouche, Bertrand Legeret, Goetz Parsiegla, Damien Sorigué, Fred Beisson, Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Environnement, Bioénergie, Microalgues et Plantes (EBMP), Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) (BIAM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Bioénergie et Microalgues (EBM), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
congenital, hereditary, and neonatal diseases and abnormalities, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Photoenzyme, Carboxy-Lyases, Phospholipid, Palmitic Acid, Chlorella, 7. Clean energy, 01 natural sciences, Biochemistry, Micelle, 03 medical and health sciences, chemistry.chemical_compound, Biofuel, Animals, Microemulsion, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], neoplasms, Micelles, Unilamellar Liposomes, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, Liposome, biology, 010405 organic chemistry, Algal Proteins, beta-Cyclodextrins, Active site, Fatty acid, Substrate (chemistry), Water, Serum Albumin, Bovine, Lipids, digestive system diseases, Hydrocarbons, 0104 chemical sciences, Kinetics, Membrane, chemistry, biology.protein, Biophysics, Biocatalysis, Cattle, Emulsions, Adsorption
Abstract

International audience; Fatty Acid Photodecarboxylase (FAP), one of the few natural photoenzymes characterized so far, is a promising biocatalyst for lipid-to-hydrocarbon conversion using light. However, the optimum supramolecular organization under which the fatty acid (FA) substrate should be presented to FAP has not been addressed. Using palmitic acid embedded in phospholipid liposomes, phospholipid-stabilized microemulsions and mixed micelles, we show that FAP displays a preference for FAs present in liposomes and at the surface of microemulsions. Adsorption kinetics onto phospholipid and galactolipid monomolecular films further suggests the ability of FAP to bind to and penetrate into membranes, with higher affinity in the presence of FAs. FAP structure reveals a potential interfacial recognition site with clusters of hydrophobic and basic residues surrounding the active site entrance. The resulting dipolar moment suggests the orientation of FAP at negatively charged interfaces. These findings provide important clues for the mode of action of FAP and the development of FAP-based bioconversion processes.

Published
2021
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27. Modulation of gastric lipase adsorption into mixed galactolipid-DPPC films by the addition of phytosterols

Author

Jeanne Kergomard, Frédéric Carrière, Gilles Paboeuf, Franck Artzner, Nathalie Barouh, Pierre Villeneuve, Claire Bourlieu, Véronique Vié, Institut de Physique de Rennes (IPR), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS), Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Bioénergétique et Ingénierie des Protéines (BIP ), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Démarche intégrée pour l'obtention d'aliments de qualité (UMR QualiSud), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Avignon Université (AU)-Université de La Réunion (UR)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro - Montpellier SupAgro, Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Avignon Université (AU)-Université de La Réunion (UR)-Université de Montpellier (UM)-Institut Agro - Montpellier SupAgro, and Kergomard, Jeanne

Subjects
[PHYS]Physics [physics], [SDV.AEN] Life Sciences [q-bio]/Food and Nutrition, [PHYS.COND]Physics [physics]/Condensed Matter [cond-mat], [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, [PHYS.COND] Physics [physics]/Condensed Matter [cond-mat], ComputingMilieux_MISCELLANEOUS, [PHYS] Physics [physics]
Abstract

National audience

Published
2021

28. Quantitative monitoring of galactolipid hydrolysis by pancreatic lipase-related protein 2 using thin layer chromatography and thymol-sulfuric acid derivatization

Author

Frédéric Carrière, Jean-David Rodier, Jérôme Lecomte, Pierre Villeneuve, Moulay Sahaka, Dominique Lafont, Brigitte Gontero, Sawsan Amara, Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Lipolytech, Département Performances des systèmes de production et de transformation tropicaux (Cirad-PERSYST), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro - Montpellier SupAgro, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), Démarche intégrée pour l'obtention d'aliments de qualité (UMR QualiSud), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Avignon Université (AU)-Université de La Réunion (UR)-Université de Montpellier (UM)-Institut Agro - Montpellier SupAgro, Gattefossé, Institut de Chimie et Biochimie Moléculaires et Supramoléculaires (ICBMS), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-École Supérieure de Chimie Physique Électronique de Lyon (CPE)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Agence Nationale de la Recherche, Aix-Marseille Université, Centre National de la Recherche Scientifique, ANR-09-CP2D-0006,GALACTOLIPASE,Utilisation de galactolipases pour extraire les acides gras de la principale source de lipides présente sur la terre. - Using galactolipases for extracting fatty acids from the main source of lipids on the earth.(2009), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Avignon Université (AU)-Université de La Réunion (UR)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), and Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS)

Subjects
L51 - Physiologie animale - Nutrition, Carbohydrate, Galactolipid, Clinical Biochemistry, 030226 pharmacology & pharmacy, 01 natural sciences, Biochemistry, Intestinal absorption, Analytical Chemistry, 03 medical and health sciences, Hydrolysis, chemistry.chemical_compound, 0302 clinical medicine, Analyse quantitative, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, L50 - Physiologie et biochimie animales, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Derivatization, Galactolipase activity, Dietary lipids, gastrointestinal lipolysis, Chromatography, Lipolyse, Chemistry, Chromatographie en couche mince, 010401 analytical chemistry, Galactose, Galactolipids, Glycolipide, galactolipase, Cell Biology, General Medicine, Thin-layer chromatography, 0104 chemical sciences, Acide gras, enzyme, Reagent, Digestion, Glycolipids, Réactif chimique, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition
Abstract

International audience; Galactolipids are the most abundant lipids on earth where they are mainly found in photosynthetic membranes of plant, algae, and cyanobacteria. Pancreatic lipase-related protein 2 (PLRP2) is an enzyme with galactolipase activity allowing mammals, especially herbivores, to digest this important source of fatty acids. We present a method for the quantitative analysis of galactolipids and galactosylated products resulting from their digestion by guinea pig PLRP2 (GPLRP2), using thin-layer-chromatography (TLC), thymolsulfuric acid as derivatization reagent and scanning densitometry for detection. Thymolsulfuric acid reagent has been used for the colorimetric detection of carbohydrates. It is shown here that the derivatization of galactosyl group from galactolipids by this reagent is not affected by the bound acyl glycerol, acyl chains length and number of galactose residues in the polar head. This allowed quantifying simultaneously the initial substrate and all galactosylated products generated upon the hydrolysis of monogalactosyl di-octanoylglycerol (C8-MGDG) by GPLRP2 using a single calibration with C8-MGDG as reference standard. The reaction products, monogalactosyl monooctanoyl glycerol (C8-MGMG) and monogalactosyl glycerol (MGG), were identified and quantified, MGG being recovered from the aqueous and analyzed by a separate TLC analysis. This method is therefore suitable to quantify the products resulting from the release of both fatty acids present in MGDG and thereby shows that PLRP2 can contribute to the complete digestion of galactolipids and further intestinal absorption of their fatty acids.

Published
2021
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29. Natural deep eutectic solvents: Hypothesis for their possible roles in cellular functions and interaction with membranes and other organized biological systems

Author

Erwann Durand, Pierre Villeneuve, Frédéric Carrière, Claire Bourlieu-lacanal, Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro - Montpellier SupAgro, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), Département Performances des systèmes de production et de transformation tropicaux (Cirad-PERSYST), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)

Subjects
F60 - Physiologie et biochimie végétale, [SDV]Life Sciences [q-bio], Cellular functions, Computational biology, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Membrane-less organelles, 01 natural sciences, 03 medical and health sciences, Biochemical reactions, L50 - Physiologie et biochimie animales, 030304 developmental biology, 0303 health sciences, Membranes, 010405 organic chemistry, Chemistry, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Membrane cellulaire, Cell functioning, Subcellular assemblies, Structure cellulaire, U50 - Sciences physiques et chimie, 0104 chemical sciences, NaDES, Membrane, Cell metabolism, Solvant
Abstract

International audience; The involvement of natural deep eutectic solvents (NaDES) in biological processes has been speculated but experimental evidence based on their observation in cells and biochemical reactions is still largely missing. Nevertheless, a sum of observations and analogies between NaDES physical-chemical properties and those of peculiar cellular environments allows raising interesting hypothesis on the role they could play. To contribute to the current thinking on NaDES in biological systems, an overview of these unusual liquid media is first presented with a special focus on how their occurrence in Nature (animal and plant) and hypothetical roles in cell metabolism were supported by experimental approaches. Then, a discussion pointing out the diversity of membranes formation and structures, describes, while challenging the readers, how NaDES may be related with membrane systems. The occurrence of NaDES in the cell and their possible interaction with highly organized intracellular structures and membrane-less organelles is also discussed. Observations already made with NaDES metabolites in cells allow discussing the possible roles and effects of NaDES on cell metabolism in connection with membranes or subcellular assemblies. Finally, suggestions for further investigations regarding the occurrence and the contribution of NaDES in the regulation of cellular functions are proposed.

Published
2021
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30. Enzymes Involved in Lipid Digestion

Author

Myriam M. L. Grundy, Ahmed Aloulou, Amal Salhi, Frédéric Carrière, Laboratoire de Biochimie et de Génie Enzymatique des Lipases, École Nationale d'Ingénieurs de Sfax | National School of Engineers of Sfax (ENIS), Université de Sfax - University of Sfax, Bioénergétique et Ingénierie des Protéines (BIP ), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), University of Reading (UOR), Myriam M.-L. Grundy, Peter J. Wilde, and Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
chemistry.chemical_classification, 0303 health sciences, biology, Cholesterol, Galactolipids, Small intestine, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, medicine.anatomical_structure, Enzyme, Biochemistry, chemistry, 030220 oncology & carcinogenesis, biology.protein, medicine, Gastric lipase, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Lipase, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Lipid digestion, 030304 developmental biology, Lingual lipase
Abstract

International audience; Lipid digestion is a complex process that takes place at the lipid-water interface and involves various lipolytic enzymes present predominantly in the stomach and the small intestine [34]. These enzymes catalyse the hydrolysis of a variety of dietary lipids from animal and plant sources, such as triacylglycerols (TAGs), phospholipids, galactolipids, cholesterol and vitamin esters. They include gastric lipase, colipase-dependent pancreatic lipase, pancreatic lipase-related proteins 2 (PLRP2), carboxyl ester hydrolase or bile salt-stimulated lipase (CEH, BSSL), and pancreatic phospholipase A2. A debate still exist about the existence of a lingual lipase in human [30, 86, 140], an enzyme that has been demonstrated to be present and active in rat and mice tongue only and which is the product of a gene ortholog [53] to the gene of gastric lipase [24] in humans and many other species. Bakala N’Goma et al. [12] have reviewed the key findings that support the existence of lingual or gastric lipases in several species in term of gene expression, enzyme immunocytolocalization and lipase activity. So far, no supporter of the existence of a lingual lipase in humans has been able to provide similar data.

Published
2021
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31. Homogeneous triacylglycerol tracers can impact fat crystallization and its lipolysis rate under simulated physiological conditions

Author

Frédéric Carrière, Cécile Vors, Marie-Caroline Michalski, Carole Knibbe, Sabine Danthine, Annie Durand, Marion Létisse, Gembloux Agro-Bio Tech [Gembloux], Université de Liège, Cardiovasculaire, métabolisme, diabétologie et nutrition (CarMeN), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Hospices Civils de Lyon (HCL), Bioénergétique et Ingénierie des Protéines (BIP ), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Artificial Evolution and Computational Biology (BEAGLE), Laboratoire de Biométrie et Biologie Evolutive - UMR 5558 (LBBE), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Inria Grenoble - Rhône-Alpes, Institut National de Recherche en Informatique et en Automatique (Inria)-Laboratoire d'InfoRmatique en Image et Systèmes d'information (LIRIS), Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-École Centrale de Lyon (ECL), Université de Lyon-Université Lumière - Lyon 2 (UL2)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-École Centrale de Lyon (ECL), Université de Lyon-Université Lumière - Lyon 2 (UL2), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'InfoRmatique en Image et Systèmes d'information (LIRIS), Université Lumière - Lyon 2 (UL2)-École Centrale de Lyon (ECL), Université de Lyon-Université de Lyon-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université Lumière - Lyon 2 (UL2)-École Centrale de Lyon (ECL), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Inria Grenoble - Rhône-Alpes, Institut National de Recherche en Informatique et en Automatique (Inria)-Institut National de Recherche en Informatique et en Automatique (Inria)-Laboratoire de Biométrie et Biologie Evolutive - UMR 5558 (LBBE), and Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Homogeneous, law, Chemistry, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], [SDV]Life Sciences [q-bio], Biophysics, [CHIM.CRIS]Chemical Sciences/Cristallography, Lipolysis, [PHYS.PHYS.PHYS-CHEM-PH]Physics [physics]/Physics [physics]/Chemical Physics [physics.chem-ph], Crystallization, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, ComputingMilieux_MISCELLANEOUS, law.invention
Abstract

International audience

Published
2020

32. The endosomal lipid bis(monoacylglycero) phosphate as a potential key player in the mechanism of action of chloroquine against SARS-COV-2 and other enveloped viruses hijacking the endocytic pathway

Author

Michel Record, Frédéric Carrière, Sonia Longhi, Centre National de la Recherche Scientifique (CNRS), Bioénergétique et Ingénierie des Protéines (BIP ), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Architecture et fonction des macromolécules biologiques (AFMB), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Departement /u563 : Oncogenèse, Signalisation et Innovation thérapeutique, Centre de Physiopathologie Toulouse Purpan (CPTP), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut Claudius Regaud, Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), and Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut Claudius Regaud

Subjects
0301 basic medicine, Endosome, [SDV]Life Sciences [q-bio], viruses, Endocytic cycle, Endosomes, Dengue virus, medicine.disease_cause, Biochemistry, Antiviral Agents, Virus, Article, lipids, 03 medical and health sciences, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Viral entry, Lysosome, medicine, Humans, endosome, Phospholipidosis, [SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases, 030102 biochemistry & molecular biology, Chemistry, SARS-CoV-2, COVID-19, Chloroquine, General Medicine, Virology, Endocytosis, 3. Good health, 030104 developmental biology, medicine.anatomical_structure, lysosome, Monoglycerides, Phospholipase inhibitor, Lysophospholipids, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract

The anti-malarial drug Chloroquine (CQ) and its derivative hydroxychloroquine have shown antiviral activities in vitro against many viruses, including coronaviruses, dengue virus and the biosafety level 4 Nipah and Hendra paramyxoviruses. The in vivo efficacy of CQ in the treatment of COVID-19 is currently a matter of debate. CQ is a lysosomotrophic compound that accumulates in lysosomes, as well as in food vacuoles of Plasmodium falciparum. In the treatment of malaria, CQ impairs the digestion and growth of the parasite by increasing the pH of the food vacuole. Similarly, it is assumed that the antiviral effects of CQ results from the increase of lysosome pH and the inhibition of acidic proteases involved in the maturation of virus fusion protein. CQ has however other effects, among which phospholipidosis, characterized by the accumulation of multivesicular bodies within the cell. The increase in phospholipid species particularly concerns bis(monoacylglycero)phosphate (BMP), a specific lipid of late endosomes involved in vesicular trafficking and pH-dependent vesicle budding. It was shown previously that drugs like progesterone, the cationic amphiphile U18666A and the phospholipase inhibitor methyl arachidonyl fluoro phosphonate (MAFP) induce the accumulation of BMP in THP-1 cells and decrease cell infection by human immunodeficiency virus. HIV viral particles were found to be retained into large endosomal-type vesicles, preventing virus spreading. Since BMP was also reported to favour virus entry through hijacking of the endocytic pathway, we propose here that BMP could play a dual role in viral infection, with its antiviral effects triggered by lysosomotropic drugs like CQ., Graphical abstract Image 1, Highlights • A known effect of Chloroquine (CQ) is phospholipidosis. • The intracellular accumulation of phospholipids is marked by a large increase in BMP, an endosomal specific-lipid. • Other antiviral drugs like progesterone induce BMP accumulation and viral particles sequestration in the cell. • BMP could be associated with the antiviral effects of CQ, at least in vitro.

Published
2020
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33. A standardised semi-dynamic in vitro digestion method suitable for food – an international consensus

Author

Ana-Isabel Mulet-Cabero, Lotti Egger, Reto Portmann, Olivia Ménard, Sébastien Marze, Mans Minekus, Steven Le Feunteun, Anwesha Sarkar, Myriam M.-L. Grundy, Frédéric Carrière, Matt Golding, Didier Dupont, Isidra Recio, André Brodkorb, Alan Mackie, Norwich Research Park, Institute of Food Research [Norwich], Biotechnology and Biological Sciences Research Council (BBSRC)-Biotechnology and Biological Sciences Research Council (BBSRC), Agroscope, Science et Technologie du Lait et de l'Oeuf (STLO), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-INSTITUT AGRO Agrocampus Ouest, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), Unité de recherche sur les Biopolymères, Interactions Assemblages (BIA), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Triskelion, University of Leeds, CENTRE FOR AGRI ENVIRONMENTAL RESEARCH UNIVERSITY OF READING GBR, Partenaires IRSTEA, Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA)-Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Riddet Institute, Instituto de Investigación en Ciencias de la Alimentación CIAL, CSIC-UAM, CEI UAM+CSIC (CIAL, CSIC-UAM, CEI UAM+CSIC), Teagasc, Moorepark Food Research Centre, INFOGEST COSTAction FA1005 and subsequently the INFOGEST network funded by INRA, AGROCAMPUS OUEST, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), School of Food Science and Nutrition, University of Leeds, and European Commission

Subjects
0301 basic medicine, Consensus, Gastric Juice, 030109 nutrition & dietetics, in vitro digestion, nutriment, [SDV]Life Sciences [q-bio], Equipment Design, General Medicine, santé humaine, modèle, Models, Biological, Gastrointestinal Tract, Kinetics, 03 medical and health sciences, 030104 developmental biology, nutrition, Food Technology, Humans, Digestion, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, semi dynamic in vitro digestion method, Food Science
Abstract

The link between food and human health is increasingly a topic of interest. One avenue of study has been to assess food disintegration and interactions within the gastrointestinal tract. In vitro digestion models have been widely used to overcome the constrictions associated with in vivo methodology. The COST Action INFOGEST developed an international, harmonised protocol for static simulation of digestion in the upper gastrointestinal tract of adults. This protocol is widely used; however, it is restricted to providing end-point assessment without considering the possible structural changes. On the other hand, there are dynamic models that provide more physiologically relevant data but are expensive and difficult to access. There is a gap between these models. The method outlined in this article provides an intermediate model; it builds upon the harmonised static model and now includes crucial kinetic aspects associated with the gastric phase of digestion, including gradual acidification, fluid and enzyme secretion and emptying. This paper provides guidance and standardised recommendations of a physiologically relevant semi-dynamic in vitro simulation of upper gastrointestinal tract digestion, with particular focus on the gastric phase. Adaptations of this model have already been used to provide kinetic data on nutrient digestion and structural changes during the gastric phase that impact on nutrient absorption. Moreover, it provides a simple tool that can be used in a wide range of laboratories., The authors would like to acknowledge the INFOGEST COST Action FA1005 and subsequently the INFOGEST network funded by INRA for enabling the networking required to develop the model described in this article.

Published
2020
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34. Physico-chemical behaviors of human and bovine milk membrane extracts and their influence on gastric lipase adsorption

Author

Stéphane Pezennec, Amélie Deglaire, Olivia Ménard, Saïd Bouhallab, Didier Dupont, Eric Gicquel, Wafa Mahdoueni, Gilles Paboeuf, Frédéric Carrière, Véronique Vié, Claire Bourlieu, Science et Technologie du Lait et de l'Oeuf (STLO), Institut National de la Recherche Agronomique (INRA)-AGROCAMPUS OUEST, Institut de Physique de Rennes (IPR), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS), Synthèse Caractérisation Analyse de la Matière (ScanMAT), Enzymologie interfaciale et de physiologie de la lipolyse (EIPL), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Département Caractérisation et Élaboration des Produits Issus de l’Agriculture, Institut National de la Recherche Agronomique, Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-INSTITUT AGRO Agrocampus Ouest, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro - Montpellier SupAgro, Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Université de Rennes (UR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Milk fat globules, Lipid Bilayers, Biochemistry, Infant nutrition, chemistry.chemical_compound, chemistry.chemical_classification, [PHYS]Physics [physics], Stomach, General Medicine, Infant Formula, Sphingomyelins, Cholesterol, Milk, Membrane, Fatty Acids, Unsaturated, AFM, Human milk membrane, Bovine milk, Surface Properties, Gastric lipase adsorption, Glycerophospholipids, Complex Mixtures, 03 medical and health sciences, Adsorption, Species Specificity, Pressure, Animals, Humans, [CHIM]Chemical Sciences, Gastric lipase, Reactivity (chemistry), Triglycerides, Glycoproteins, Phosphatidylethanolamine, Chromatography, Milk, Human, 030102 biochemistry & molecular biology, Phosphatidylethanolamines, Monolayer, Infant, Lipase, Lipid Droplets, Bovine milk membrane, 030104 developmental biology, Enzyme, chemistry, Cattle, Glycolipids
Abstract

International audience; Milk fat globule membrane conditions the reactivity and enzymatic susceptibility of milk lipids. The use of bovine membrane extracts to make infant formulas more biomimetic of human milk has been suggested recently. A comparison of the physico-chemical behavior of human and bovine milk membrane extracts and their interaction with gastric lipase is here undertaken using biophysical tools. Milk membrane extracts (70% of polar lipids) were obtained either pooling of mature human milk (n = 5) or bovine buttermilk. Human extract contained more anionic glycerophospholipids, less phosphatidylethanolamine and more unsaturated fatty acids (57% versus 46%) than bovine extract. Human extract presented a higher compressibility, with slower increase of surface pressure, than bovine extract. Micronic liquid condensed (LC) domains were evidenced in both extracts at 10 mN/m, but the evolution differs upon compression. Upon gastric lipase addition, an adsorption preference for liquid expanded phase (LE) was observed for both extracts. However, insertion was more homogeneous in terms of height level in human extract and impacted less its lipid lateral organization than in bovine extract. Both membrane extracts share close physico-chemical properties, however human membrane higher compressibility may favour gastric lipase insertion and higher interfacial reactivity in gastric conditions.

Published
2020
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35. Biogenesis and fate of lipid droplets

Author

Frédéric Carrière, Yonghua Li-Beisson, Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) (BIAM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Environnement, Bioénergie, Microalgues et Plantes (EBMP), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Bioénergie et Microalgues (EBM), and Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)

Subjects
0106 biological sciences, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, [SDV]Life Sciences [q-bio], Saccharomyces cerevisiae, 01 natural sciences, Biochemistry, 03 medical and health sciences, Diabetes mellitus genetics, Mice, Lipid droplet, Diabetes Mellitus, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Obesity, 030304 developmental biology, Diatoms, 0303 health sciences, Chemistry, General Medicine, Lipid Droplets, Plants, Lipid Metabolism, Cell biology, Biofuels, lipids (amino acids, peptides, and proteins), Biogenesis, 010606 plant biology & botany
Abstract

International audience; Editorial Special issue "Biogenesis and Fate of Lipid Droplets" Intracellular lipid droplets (LDs) are receiving increasing interest from the scientific community, notably because of their link with metabolic diseases (obesity, diabetes) and the production of lipid-derived biofuels in microorganisms. Understanding the biogenesis and fate of LDs upon energy mobilization is essential to monitor fat storage in adipose tissue, but also to improve lipid productivity in microalgae under various conditions of growth [1]. Intracellular LDs are today considered as individual organelles since they host specific metabolic functions such as the biosynthesis of triacylglycerols (TAGs) and phospholipids [2]. In addition to the enzymes involved in these pathways, several proteins have been identified as associated with LDs and their number has been increasing with the progress of proteomics. Some of these proteins presenting hydrophobic patches and a high amphiphilicity are involved in the structure of LDs and together with polar lipids they form the membrane surrounding the neutral lipid core. Some others are involved in the budding of LDs from lipid bilayers and the interactions of LDs with membranes and other organelles. These interactions are important for the transfer of fatty acids and lipid remodelling and homeostasis. The combination of lipidomics and proteomics has led to a better description of LD composition, while microscopy has allowed the visualisation of the specific location of proteins at the surface of LDs. Nevertheless, a better knowledge of the lipid-protein interactions that lead to the biogenesis and fate of LDs is still required. In this context, cellular biologists now adopt principles and methods from biophysics and physical chemistry of lipid emulsions and colloids [3]. Emulsions made of TAGs, polar lipids and proteins can serve as models for intracellular LDs. The biodiversity of LDs in animals, plants and microorganisms and their various functions in lipid storage and transport also allows the comparison of data and knowledge from various fields that will help build a global understanding of structure-function relationships. The original articles and reviews of this special issue are a collection of topics presented at the 14th GERLI Lipidomics meeting, which was be held at St Maximin-la-Sainte Baume, France, from September 30th to October 3rd, 2018, with the aim of gathering researchers from various areas with a common interest in LDs, their physical chemistry and metabolism, their association with diseases and various applications in biotechnology, pharmacology and nutrition. The two first mini-reviews are related to lipid metabolism in microalgae. Lupette et al. cover the biosynthesis of fatty acids, acyl-glycero-lipids and sterols in diatoms and show how a metabolic intermediate common to these pathways, like acetyl-CoA, can play an essential role in directing the carbon flux from acyl-lipid to sterol biosynthesis and be determinant for the balance between TAGs and sterols. Prioretti et al. investigate how the target of rapamycin (TOR) signalling pathway could be modulated to enhance TAG production in microalgae. The use of TOR inhibitors increases TAG productivity in the marine diatom Phaeodactylum tricornutum, without stopping growth, while TAG accumulation in microalgae is usually observed under stress conditions with a low growth rate. Yuan et al. report on the characterization of three diacylglycerol acyltransferases (DGATs) from oil palm (Elaeis guineensis), the main source of vegetable oil on Earth. DGATs are key enzymes for the biosynthesis of TAGs and the formation of LDs. Using the heterologous expression of these DGATs in the yeast Saccharomyces cerevisiae, they show that two of them are true DGATs, that could restore TAG synthesis in a yeast TAG-deficient mutant, while the third one is a wax ester synthase.

Published
2020
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36. Storage Compound Accumulation in Diatoms as Response to Elevated CO 2 Concentration

Author

Karen Yangüez, Erik Jensen, Frédéric Carrière, Brigitte Gontero, Bioénergétique et Ingénierie des Protéines (BIP ), and Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0106 biological sciences, 0301 basic medicine, CO2, [SDV]Life Sciences [q-bio], Biomass, chrysolaminarin, 01 natural sciences, General Biochemistry, Genetics and Molecular Biology, Article, diatoms, 03 medical and health sciences, chemistry.chemical_compound, Nutrient, Food science, Chrysolaminarin, Phaeodactylum tricornutum, lcsh:QH301-705.5, General Immunology and Microbiology, biology, fungi, biology.organism_classification, Enzyme assay, 030104 developmental biology, Diatom, chemistry, lcsh:Biology (General), Navicula, CO 2, biology.protein, Seawater, triacylglycerol, General Agricultural and Biological Sciences, 010606 plant biology & botany
Abstract

Accumulation of reserve compounds (i.e., lipids and chrysolaminarin) in diatoms depends on the environmental conditions, and is often triggered by stress conditions, such as nutrient limitation. Manipulation of CO2 supply can also be used to improve both lipids and carbohydrates accumulation. Given the high diversity among diatoms, we studied the two marine model diatoms&mdash, Thalassiosira pseudonana and Phaeodactylum tricornutum, a freshwater diatom, Asterionella formosa, and Navicula pelliculosa&mdash, found in fresh- and sea-water environments. We measured the accumulation of reserve compounds and the activity of enzymes involved in carbon metabolism in these diatoms grown at high and atmospheric CO2. We observed that biomass and lipid accumulation in cells grown at high CO2 differ among the diatoms. Lipid accumulation increased only in P. tricornutum and N. pelliculosa grown in seawater in response to elevated CO2. Moreover, accumulation of lipids was also accompanied by an increased activity of the enzymes tested. However, lipid accumulation and enzyme activity decreased in N. pelliculosa cultured in fresh water. Chrysolaminarin accumulation was also affected by CO2 concentration, however, there was no clear relation with lipids accumulation. Our results are relevant to understand better the ecological role of the environment in the diatom adaptation to CO2 and the mechanisms underpinning the production of storage compounds considering diatom diversity.

Published
2019
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37. The 1,2-o-dilauryl-rac-glycero-3-glutaric acid-(6'-methylresorufin) ester (DGGR) lipase assay in cats and dogs is not specific for pancreatic lipase

Author

Panagiotis G. Xenoulis, Evangelia M Stavroulaki, Frédéric Carrière, Jan S. Suchodolski, Jonathan A. Lidbury, Jörg M. Steiner, Sue Yee Lim, Department of Small Animal Clinical Sciences, Texas A&M University [College Station], Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)

Subjects
030213 general clinical medicine, 040301 veterinary sciences, [SDV]Life Sciences [q-bio], lipoprotein lipase, Pharmacology, Cat Diseases, 0403 veterinary science, Glutarates, 03 medical and health sciences, 0302 clinical medicine, Dogs, medicine, Animals, Dog Diseases, Lipase, Canine pancreatitis, ComputingMilieux_MISCELLANEOUS, Lipoprotein lipase, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, CATS, General Veterinary, biology, Chemistry, [SDV.BA]Life Sciences [q-bio]/Animal biology, postheparin lipase, Esters, Spec cPL, 04 agricultural and veterinary sciences, Heparin, medicine.disease, Spec fPL, 3. Good health, hepatic lipase, Pancreatitis, biology.protein, Cats, Hepatic lipase, medicine.drug, Lipoprotein
Abstract

International audience; Background: The measurement of pancreatic lipase is important for the diagnosis offeline and canine pancreatitis. Recent studies have claimed that lipase assays using the 1,2-o-dilauryl-rac-glycero-3-glutaric acid-(6’-methylresorufin) ester (DGGR) as a substrate are more specific for measuring pancreatic lipase than traditional lipase assays. However, the analytical specificity of this assay for pancreatic lipase has not been demonstrated.Objectives: We aimed to determine whether hepatic and/or lipoprotein lipases can interfere with the DGGR-based assay results in cats and dogs. We, therefore, compared plasma lipase activities measured using DGGR-based and pancreatic lipase immunoreactivity (PLI) assays before and after administering heparin, known to cause the release of hepatic and lipoprotein lipases, in cats and dogs.Methods: Heparin was administered in six cats and six dogs. Blood was collected at baseline and 10, 20, 30, 60, and 120 minutes after heparin administration. Lipase activity was measured using a DGGR-based assay, and PLI concentrations were measured using the Spec fPL and cPL assays for cats and dogs, respectively.Results: Plasma lipase activities, as measured using the DGGR-based assay, increased significantly 10 minutes after heparin administration in both cats (P = .003) and dogs (P = .006) and returned to baseline by 120 minutes. In contrast, PLI concentrations showed no significant changes after heparin administration.Conclusions: DGGR is not only hydrolyzed by pancreatic lipase but also by hepatic lipase, lipoprotein lipase, or both, in cats and dogs. Since these extrapancreatic lipases are also naturally present in cats and dogs, they could contribute to the lack of analytical specificity for the DGGR-based assays.

Published
2019
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38. Functional characterization and FTIR-based 3D modeling of full length and truncated forms of Scorpio maurus venom phospholipase A 2

Author

Hélène Gaussier, Hanen Louati, Frédéric Carrière, Raida Jallouli, Goetz Parsiegla, Pascal Mansuelle, Najeh Krayem, Youssef Gargouri, ENIS, Lab Biochim & Genie Enzymat Lipases, Route Soukra, BP 1173, Sfax 3038, Tunisia, Université de Sfax - University of Sfax, Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Institut méditerranéen d'océanologie (MIO), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Toulon (UTLN)-Centre National de la Recherche Scientifique (CNRS), Plateforme Protéomique [Marseille], Institut de Microbiologie de la Méditerranée (IMM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), and Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université de Toulon (UTLN)

Subjects
0301 basic medicine, Biophysics, Phospholipid, Scorpio maurus, Molecular modeling, Molecular cloning, Phospholipase, Secreted phospholipase A2, Biochemistry, law.invention, 03 medical and health sciences, chemistry.chemical_compound, Phospholipase A2, law, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Enzyme activity, Molecular Biology, Phospholipase A, 030102 biochemistry & molecular biology, biology, Chemistry, biology.organism_classification, Secreted phospholipase A 2, FTIR spectroscopy, 030104 developmental biology, biology.protein, Recombinant DNA, Heterologous expression
Abstract

International audience; Background: Heterodimeric phospholipase A2 from venom glands of Tunisian scorpion Scorpio maurus (Sm-PLGV) had been purified. It contains long and short chains linked by a disulfide bridge. Sm-PLGV exhibits hemolytic activity towards human erythrocytes and interacts with phospholipid monolayers at high surface pressure. The investigation of structure-function relationships should provide new clues to understand its activity.Methods: Molecular cloning of Sm-PLGV and heterologous expression in Escherichia coli of three recombinant forms was used to determine the role of the short chain on enzymatic activity. Infrared spectroscopy assisted 3D model building of the three recombinant constructs (phospholipases with and without the penta-peptide and Long chain only) allowed us to propose an explanation of the differences in specific activities and their interaction with various phospholipids.Results: Nucleotide sequence of Sm-PLGV encodes 129 residues corresponding to the Long chain, the penta-peptide and the short chain. Although recombinant phospholipases without and with the penta-peptide have different specific activities, they display a similar substrate specificity on various phospholipid monolayers and similar bell-shaped activity profiles with maxima at high surface pressure. The absence of the short chain reduces significantly enzymatic and hemolytic activities. The 3D models pointed to an interaction of the short chain with the catalytic residues, what might explain the difference in activities of our constructs.Conclusion: Infrared spectroscopy data and 3D modeling confirm the experimental findings that highlight the importance of the short chain for the Sm-PLGV activity.

Published
2018
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39. IR spectroscopy analysis of pancreatic lipase-related protein 2 interaction with phospholipids: 1. Discriminative recognition of mixed micelles versus liposomes

Author

Sylvie Robert, Jean-Claude Bakala N’Goma, Frédéric Carrière, Eduardo Mateos-Diaz, Deborah Byrne, Hélène Gaussier, Institut de Microbiologie de la Méditerranée (IMM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Enzymologie interfaciale et de physiologie de la lipolyse (EIPL), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Bioénergétique et Ingénierie des Protéines (BIP ), Institut méditerranéen d'océanologie (MIO), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Toulon (UTLN)-Centre National de la Recherche Scientifique (CNRS), and TADEO, DANIELE

Subjects
0301 basic medicine, Spectrophotometry, Infrared, Guinea Pigs, Phospholipid, Infrared spectroscopy, Biochemistry, Micelle, lipids, 03 medical and health sciences, chemistry.chemical_compound, Dynamic light scattering, Phosphatidylcholine, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Animals, Organic chemistry, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, phospholipase, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Pancreas, Molecular Biology, Micelles, Phospholipids, ComputingMilieux_MISCELLANEOUS, lipid digestion, Liposome, 030102 biochemistry & molecular biology, Hydrolysis, Vesicle, Organic Chemistry, technology, industry, and agriculture, Lipase, Cell Biology, [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, enzyme, FTIR spectroscopy, Crystallography, 030104 developmental biology, chemistry, Liposomes, lipolysis, lipids (amino acids, peptides, and proteins), Lipid digestion
Abstract

International audience; Guinea pig pancreatic lipase-related protein 2 (GPLRP2) is an interesting model enzyme that can hydrolyze a large set of acylglycerols in vitro but displays however some selectivity depending on the supramolecular structure of substrate and the presence of surfactants like bile salts. We showed that GPLRP2 hydrolyzes 1,2-Dipalmitoyl phosphatidylcholine (DPPC) present in mixed micelles with sodium taurodeoxycholate (NaTDC) but not in multilamellar (MLV) and large unilamellar (LUV) vesicles of DPPC. After characterization of these lipid aggregates by dynamic light scattering (DLS), the discriminative recognition of DPPC in DPPC/NaTDC micelles versus MLV and LUV by an inactive variant (S152G) of GPLRP2 to avoid the effect of substrate hydrolysis was investigated using Fourier transform infrared spectroscopy (FTIR). IR spectra were recorded after hydrogen/deuterium exchange, at pD 6 and various temperatures to study phase transitions. We analyzed the methylene asymmetric stretching ((CH2)as), the carbonyl stretching ((CO)) and the composite polar head-group vibration bands, first to characterized differences in DPPC micelles and vesicles, and second to estimate the degree of interaction of GPLRP2 S152G with phospholipid. Our results indicate that a significant interaction between GPLRP2 S152G and DPPC is only observed when NaTDC is added to the system to form micelles and this can be explained by the different organization of DPPC in mixed micelles compared to lamellar vesicles (higher hydration of polar head, higher mobility of alkyl chains) that favors GPLRP2 penetration into the phospholipid layer.

Published
2018
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40. IR spectroscopy analysis of pancreatic lipase-related protein 2 interaction with phospholipids: 2. Discriminative recognition of various micellar systems and characterization of PLRP2-DPPC-bile salt complexes

Author

Hélène Gaussier, Frédéric Carrière, Deborah Byrne, Eduardo Mateos-Diaz, Moulay Sahaka, Priscila Sutto-Ortiz, Enzymologie interfaciale et de physiologie de la lipolyse (EIPL), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Institut de Microbiologie de la Méditerranée (IMM), Institut méditerranéen d'océanologie (MIO), and Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Toulon (UTLN)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, 1,2-Dipalmitoylphosphatidylcholine, Spectrophotometry, Infrared, [SDV]Life Sciences [q-bio], Guinea Pigs, Size-exclusion chromatography, Salt (chemistry), Infrared spectroscopy, Phospholipase, Biochemistry, Micelle, Bile Acids and Salts, lipids, 03 medical and health sciences, chemistry.chemical_compound, Hydrolysis, Polymer chemistry, Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, phospholipase, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Pancreas, Molecular Biology, Micelles, lipid digestion, chemistry.chemical_classification, Chromatography, 030102 biochemistry & molecular biology, Organic Chemistry, technology, industry, and agriculture, Lipase, Cell Biology, [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, enzyme, FTIR spectroscopy, 030104 developmental biology, Monomer, chemistry, lipolysis, lipids (amino acids, peptides, and proteins), Lipid digestion
Abstract

International audience; The interaction of pancreatic lipase-related protein 2 (PLRP2) with various micelles containing phospholipids was investigated using pHstat enzyme activity measurements, differential light scattering, size exclusion chromatography (SEC) and transmission IR spectroscopy. Various micelles of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and lysophosphatidylcholine were prepared with either bile salts (sodium taurodeoxycholate or glycodeoxycholate) or Triton X-100, which are substrate-dispersing agents commonly used for measuring phospholipase activities. PLRP2 displayed a high activity on all phospholipid-bile salt micelles, but was totally inactive on phospholipid-Triton X-100 micelles. These findings clearly differentiate PLRP2 from secreted pancreatic phospholipase A2 which is highly active on both types of micelles. Using an inactive variant of PLRP2, SEC experiments allowed identifying two populations of PLRP2-DPPC-bile salt complexes corresponding to a high molecular weight 1:1 PLRP2-micelle association and to a low molecular weight association of PLRP2 with few monomers of DPPC / bile salts. IR spectroscopy analysis showed how DPPC-bile salt micelles differ from DPPC-Triton X-100 micelles by a higher fluidity of acyl chains and higher hydration / H-bonding of the interfacial carbonyl region. The presence of bile salts allowed observing changes in the IR spectrum of DPPC upon addition of PLRP2 (higher rigidity of acyl chains, dehydration of the interfacial carbonyl region), while no change was observed with Triton X-100. The differences between these surfactants and their impact on substrate recognition by PLRP2 are discussed, as well as the mechanism by which high and low molecular weight PLRP2-DPPC-bile salt complexes may be involved in the overall process of DPPC hydrolysis.

Published
2018
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41. Evaluation of vitamin D bioaccessibility and mineral solubility from test meals containing meat and/or cereals and/or pulses using in vitro digestion

Author

Charlotte Halimi, Giulia Scorrano, Tiffany Antoine, Christèle Icard-Vernière, Stéphane Georgé, Claire Mouquet-Rivier, Frédéric Carrière, Emmanuelle Reboul, Amal Salhi, Centre recherche en CardioVasculaire et Nutrition = Center for CardioVascular and Nutrition research (C2VN), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Nutrition et Alimentation des Populations aux Suds (NutriPass), Université Montpellier 1 (UM1)-Institut de Recherche pour le Développement (IRD)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Centre Technique de la Conservation des Produits Agricoles, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Démarche intégrée pour l'obtention d'aliments de qualité (UMR QualiSud), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Avignon Université (AU)-Université de La Réunion (UR)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut de Recherche pour le Développement (IRD)-Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Université de Montpellier (UM), and CCSD, Accord Elsevier

Subjects
Vitamin, Meat, Bioavailability, Iron, Lipolysis, Biological Availability, chemistry.chemical_element, Zinc, 01 natural sciences, Analytical Chemistry, chemistry.chemical_compound, 0404 agricultural biotechnology, Vitamin D and neurology, Humans, Pancreatic lipase, Magnesium, Food science, Vitamin D, Solubility, Meals, Cholecalciferol, 2. Zero hunger, Minerals, digestive, oral, and skin physiology, 010401 analytical chemistry, Chickpeas, 04 agricultural and veterinary sciences, General Medicine, Micronutrient, 040401 food science, 0104 chemical sciences, [SDV.AEN] Life Sciences [q-bio]/Food and Nutrition, chemistry, Digestion, Edible Grain, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Food Science
Abstract

International audience; In this study, we evaluated vitamin D and mineral (iron, zinc, magnesium) transfer to the bolus aqueous phase during the digestion of meals with/without pulses. We performed in vitro digestions using test meals made either of i) beef and/or semolina and/or chickpeas, or of ii) potatoes supplemented or not with fibers, phytates, tannins and saponins. Chickpea presence led to a decrease in vitamin D bioaccessibility (-56%, p < 0.05) and mineral solubility (-28% for iron, p < 0.05) compared with meals with beef and/or semolina only. This effect was largely compensated for vitamin D by the fact that this vitamin was more stable during digestion of meals based on plant foods only than of meals with beef. Tannins were the most deleterious compounds for iron solubility, while phytates and tannins decreased vitamin D bioaccessibility. Agronomical or technical solutions to selectively decrease the amount in pulses of compounds that affect micronutrient bioavailability should be further explored.

Published
2021
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42. Efficient heterologous expression of Fusarium solani lipase, FSL2, in Pichia pastoris, functional characterization of the recombinant enzyme and molecular modeling

Author

Raida Jallouli, Youssef Gargouri, Goetz Parsiegla, Frédéric Carrière, and Sofiane Bezzine

Subjects
0301 basic medicine, Gene Expression, Biochemistry, Pichia, Substrate Specificity, Pichia pastoris, Fungal Proteins, 03 medical and health sciences, Fusarium, Structural Biology, Catalytic Domain, Complementary DNA, Catalytic triad, Cloning, Molecular, Lipase, Molecular Biology, biology, Molecular mass, Active site, General Medicine, Hydrogen-Ion Concentration, biology.organism_classification, Recombinant Proteins, Molecular Docking Simulation, 030104 developmental biology, Structural Homology, Protein, biology.protein, Calcium, Heterologous expression, Fusarium solani, Protein Binding
Abstract

The gene coding for a lipase of Fusarium solani, designated as FSL2, shows an open reading frame of 906bp encoding a 301-amino acid polypeptide with a molecular mass of 30kDa. Based on sequence similarity with other fungal lipases, FSL2 contains a catalytic triad, consisting of Ser144, Asp198, and His256. FSL2 cDNA was subcloned into the pGAPZαA vector containing the Saccharomyces cerevisiae α-factor signal sequence and this construct was used to transform Pichia pastoris and achieve a high-level extracellular production of a FSL2 lipase. Maximum lipase activity was observed after 48h. The optimum activity of the purified recombinant enzyme was measured at pH 8.0-9.0 and 37°C. FSL2 is remarkably stable at alkaline pH values up to 12 and at temperatures below 40°C. It has high catalytic efficiency towards triglycerides with short to long chain fatty acids but with a marked preference for medium and long chain fatty acids. FSL2 activity is decreased at sodium taurodeoxycholate concentrations above the Critical Micelle Concentration (CMC) of this anionic detergent. However, lipase activity is enhanced by Ca2+ and inhibited by EDTA or Cu2+ and partially by Mg2+ or K+. In silico docking of medium chain triglycerides, monogalctolipids (MGDG), digalactolipids (DGDG) and long chain phospholipids in the active site of FSL2 reveals structural solutions.

Published
2017
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43. Orchestration of algal metabolism by protein disorder

Author

Hélène Launay, Frédéric Carrière, Véronique Receveur-Bréchot, Brigitte Gontero, Bioénergétique et Ingénierie des Protéines (BIP ), and Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, [SDV]Life Sciences [q-bio], Biophysics, RuBisCo, Chaperone, Intrinsically disordered proteins, EPYC, Biochemistry, Pyrenoid, 03 medical and health sciences, Chlorophyta, Heat shock protein, Microalgae, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Photosynthesis, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Molecular Biology, 030102 biochemistry & molecular biology, biology, Chemistry, RuBisCO, Algal Proteins, CP12, Hsp90, Cell biology, Intrinsically Disordered Proteins, [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, 030104 developmental biology, Proteostasis, Calvin cycle, Chaperone (protein), Hsp33, biology.protein, Molecular Chaperones
Abstract

International audience; Intrinsically disordered proteins (IDPs) are proteins that provide many functional advantages in a large number of metabolic and signalling pathways. Because of their high flexibility that endows them with pressure-, heat- and acid-resistance, IDPs are valuable metabolic regulators that help algae to cope with extreme conditions of pH, temperature, pressure and light. They have, however, been overlooked in these organisms. In this review, we present some well-known algal IDPs, including the conditionally disordered CP12, a protein involved in the regulation of CO2 assimilation, as probably the best known example, whose disorder content is strongly dependent on the redox conditions, and the essential pyrenoid component 1 that serves as a scaffold for ribulose-1, 5-bisphosphate carboxylase/oxygenase. We also describe how some enzymes are regulated by protein regions, called intrinsically disordered regions (IDRs), such as ribulose-1, 5-bisphosphate carboxylase/oxygenase activase, the A2B2 form of glyceraldehyde-3-phosphate dehydrogenase and the adenylate kinase. Several molecular chaperones, which are crucial for cell proteostasis, also display significant disorder propensities such as the algal heat shock proteins HSP33, HSP70 and HSP90. This review confirms the wide distribution of IDPs in algae but highlights that further studies are needed to uncover their full role in orchestrating algal metabolism.

Published
2019
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44. Studies of crab digestive phospholipase acting on phospholipid monolayers: Activation by temperature

Author

Ahmed Fendri, Adel Sayari, Slim Cherif, Frédéric Carrière, Faculté des Sciences de Gafsa, Université de Gafsa, Institut Supérieur de Biotechnologie de Sfax (ISBS), Université de Sfax - University of Sfax, Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), École Nationale d'Ingénieurs de Sfax | National School of Engineers of Sfax (ENIS), and Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)

Subjects
Brachyura, Surface Properties, Swine, Phospholipid, 02 engineering and technology, Phospholipase, Biochemistry, Catalysis, Phase Transition, 03 medical and health sciences, Hydrolysis, chemistry.chemical_compound, Membrane Lipids, Structural Biology, Bromide, Monolayer, Molecule, Animals, Transition Temperature, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Phospholipases A2, Secretory, Molecular Biology, Phospholipids, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, Chemistry, General Medicine, 021001 nanoscience & nanotechnology, [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, Kinetics, Enzyme, Phospholipases, Biophysics, lipids (amino acids, peptides, and proteins), Digestion, 0210 nano-technology
Abstract

International audience; The water-soluble lipolytic enzymes act at the interface of insoluble lipid substrates, where the catalytical step is coupled with various interfacial phenomena as enzyme penetration, solubilization of reaction products, loss of mechanical stability of organized assemblies of phospholipids molecule, etc. Using the classical emulsified system and the monomolecular film technique, we compared the interfacial properties of crab digestive phospholipase (CDPL) with those of the porcine pancreatic one (PPPL). A kinetic study on the surface pressure dependency of the two phospholipases was performed using monomolecular films of three substrates: di C12-PC (1.2-dilauroyl-sn-glycerol-3-phosphocholine); di C12-PG (1.2-dilauroyl-sn-glycerol phosphoglycerol) and di C12-PE (1.2- dilauroyl-sn-glycerol phosphoethanolamine). The use of a substrate in monofilm state allows the monitoring, during the biocatalysed reactions, of several physico-chemical parameters and permits the modification of the “quality of interface”. The effect of the temperature on the hydrolysis rate of these substrates was also checked. Our results show that specific activities of both phospholipases were affected by the variation of the subphase temperature. CDPL was irreversibly inactivated by p-bromo-phenacyl bromide, the specific inhibitor of secretory PLA2s. The hyperbolic behaviour observed was coherent with hopping mode of action, one of the two characteristic interfacial mechanisms of PLA2s.

Published
2019
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45. Characterization of all the lipolytic activities in pancreatin and comparison with porcine and human pancreatic juices

Author

Sawsan Amara, Amal Salhi, Rémy Puppo, Ahmed Aloulou, Frédéric Carrière, Régine Lebrun, Pascal Mansuelle, Brigitte Gontero, Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), École Nationale d'Ingénieurs de Sfax | National School of Engineers of Sfax (ENIS), Lipolytech, Plateforme Protéomique [Marseille], Institut de Microbiologie de la Méditerranée (IMM), and Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Swine, Pancreatic Extracts, Phospholipase, Biochemistry, Carboxylesterase, 03 medical and health sciences, Gastrointestinal Agents, Pancreatic Juice, Species Specificity, Galactolipase, Enzyme Stability, medicine, Animals, Humans, Amylase, Amino Acid Sequence, Lipase, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Pancreas, Enzyme Assays, chemistry.chemical_classification, lipid digestion, 030102 biochemistry & molecular biology, biology, Sequence Homology, Amino Acid, General Medicine, Hydrogen-Ion Concentration, Sterol Esterase, Kinetics, 030104 developmental biology, Enzyme, medicine.anatomical_structure, chemistry, Phospholipases, Pancreatin, biology.protein, Exocrine Pancreatic Insufficiency, Lipid digestion, Carboxylic Ester Hydrolases, Sequence Alignment
Abstract

International audience; Porcine pancreatic extracts (PPE), also named pancreatin, are commonly used as a global source of pancreatic enzymes for enzyme replacement therapy in patients with exocrine pancreatic insufficiency. They are considered as a good substitute of human pancreatic enzymes and they have become a material of choice for in vitro models of digestion. Nevertheless, while the global PPE contents in lipase, protease and amylase activities are well characterized, little is known about individual enzymes. Here we characterized the lipase, phospholipase, cholesterol esterase and galactolipase activities of PPE and compared them with those of porcine (PPJ) and human (HPJ) pancreatic juices. The phospholipase to lipase activity ratio was similar in PPJ and HPJ, but was 4-fold lower in PPE. The galactolipase and cholesterol esterase activities were found at lower levels in PPJ compared to HPJ, and they were further reduced in PPE. The enzymes known to display these activities in HPJ, pancreatic lipase-related protein 2 (PLRP2) and carboxylester hydrolase/bile salt-stimulated lipase (CEH/BSSL), were identified in PPJ using gel filtration experiments, SDS-PAGE and LC-MS/MS analysis. The galactolipase and cholesterol esterase activities of PPE indicated that PLRP2 and CEH/BSSL are still present at low levels in this enzyme preparation, but they were not detected by mass spectrometry. Besides differences between porcine and human enzymes, the lower levels of phospholipase, galactolipase and cholesterol esterase activities in PPE are probably due to some proteolysis occurring during the production process. In conclusion, PPE do not provide a full substitution of the lipolytic enzymes present in HPJ.

Published
2019
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46. Identification of a new natural gastric lipase inhibitor from star anise

Author

Michele Fiore, Imed Koubaa, Ahmed Aloulou, Karine Alvarez, Mohamed Sellami, Renaud Rahier, Jannet Kamoun, Pascal Mansuelle, Régine Lebrun, Abdelkarim Abousalham, Frédéric Carrière, Alexandra Berlioz-Barbier, École Nationale d'Ingénieurs de Sfax | National School of Engineers of Sfax (ENIS), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Métabolisme, Enzymes et Mécanismes Moléculaires (MEM²), Institut de Chimie et Biochimie Moléculaires et Supramoléculaires (ICBMS), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS), Faculté des Sciences de Sfax, Université de Sfax - University of Sfax, Institut de Microbiologie de la Méditerranée (IMM), Plateforme Protéomique [Marseille], Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Plateforme de Spectrométrie de Masse Protéomique - Mass Spectrometry Proteomics Platform (MSPP), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Institut National de la Recherche Agronomique (INRA), Chimie Organique 2-Glycochimie (CO2GLYCO), Architecture et fonction des macromolécules biologiques (AFMB), Institut National de la Recherche Agronomique (INRA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Enzymologie interfaciale et de physiologie de la lipolyse (EIPL), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Centre National de la Recherche Scientifique (CNRS)-École Supérieure Chimie Physique Électronique de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-École Supérieure Chimie Physique Électronique de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon, Institut National de la Recherche Agronomique (INRA)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Institut National de la Recherche Agronomique (INRA), Laboratoire de Biochimie et de Génie Enzymatique des Lipases, Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS), Algorithmes, Composants, Modèles Et Services pour l'informatique répartie (ACMES-SAMOVAR), Services répartis, Architectures, MOdélisation, Validation, Administration des Réseaux (SAMOVAR), Institut Mines-Télécom [Paris] (IMT)-Télécom SudParis (TSP)-Institut Mines-Télécom [Paris] (IMT)-Télécom SudParis (TSP), Département Informatique (INF), Institut Mines-Télécom [Paris] (IMT)-Télécom SudParis (TSP), Institut Polytechnique de Paris (IP Paris), Centre National de la Recherche Scientifique (CNRS), Centre Commun de Spectrometrie de Masse (CCSM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université de Lyon-Université de Lyon-École Supérieure de Chimie Physique Électronique de Lyon (CPE)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Département Informatique (TSP - INF), Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), and Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Star anise, [SDV]Life Sciences [q-bio], Flavonoid, Future application, Ether, Illicium, Mass Spectrometry, Serine, 03 medical and health sciences, chemistry.chemical_compound, Peptide mass fingerprinting, Gastric lipase, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Enzyme Inhibitors, Inhibition, chemistry.chemical_classification, 030109 nutrition & dietetics, Binding Sites, Chemistry, Plant Extracts, Stomach, General Medicine, Lipase, 3. Good health, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Molecular Docking Simulation, Kinetics, 030104 developmental biology, Biochemistry, Molecular docking, Lipase inhibitors, Drug delivery, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Guided fractionation, Food Science
Abstract

International audience; The identification and isolation of bioactive compounds are of great interest in the drug delivery field, despite being a difficult task. We describe here an innovative strategy for the identification of a new gastric lipase inhibitor from star anise for the treatment of obesity. After plant screening assays for gastric lipase inhibition, star anise was selected and investigated by bioactivity guided fractionation. MALDI-TOF mass spectrometry and peptide mass fingerprinting allowed the detection of an inhibitor covalently bound to the catalytic serine of gastric lipase. A mass-directed screening approach using UPLC-HRMS and accurate mass determination searching identified the flavonoid myricitrin-5-methyl ether (M5ME) as a lipase inhibitor. The inhibitory activity was rationalized based on molecular docking, showing that M5ME is susceptible to nucleophilic attack by gastric lipase. Overall, our data suggest that M5ME may be considered as a potential candidate for future application as a gastric lipase inhibitor for the treatment of obesity.

Published
2019
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47. Homogeneous triacylglycerol tracers have an impact on the thermal and structural properties of dietary fat and its lipolysis rate under simulated physiological conditions

Author

Damien Agopian, Marie-Caroline Michalski, Annie Durand, Frédéric Carrière, Marion Létisse, Sabine Danthine, Romain Guyon, Cécile Vors, Carole Knibbe, Gembloux Agro-Bio Tech [Gembloux], Université de Liège, Cardiovasculaire, métabolisme, diabétologie et nutrition (CarMeN), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Artificial Evolution and Computational Biology (BEAGLE), Laboratoire d'InfoRmatique en Image et Systèmes d'information (LIRIS), Université Lumière - Lyon 2 (UL2)-École Centrale de Lyon (ECL), Université de Lyon-Université de Lyon-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université Lumière - Lyon 2 (UL2)-École Centrale de Lyon (ECL), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Inria Grenoble - Rhône-Alpes, Institut National de Recherche en Informatique et en Automatique (Inria)-Institut National de Recherche en Informatique et en Automatique (Inria)-Laboratoire de Biométrie et Biologie Evolutive - UMR 5558 (LBBE), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Centre National Interprofessionnel de l'Economie Laitiere (CNIEL, French Dairy Interbranch Organization) INRA CNIEL, Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Laboratoire de Biométrie et Biologie Evolutive - UMR 5558 (LBBE), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Inria Grenoble - Rhône-Alpes, Institut National de Recherche en Informatique et en Automatique (Inria)-Laboratoire d'InfoRmatique en Image et Systèmes d'information (LIRIS), Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-École Centrale de Lyon (ECL), Université de Lyon-Université Lumière - Lyon 2 (UL2)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-École Centrale de Lyon (ECL), Université de Lyon-Université Lumière - Lyon 2 (UL2), Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Lumière - Lyon 2 (UL2)-École Centrale de Lyon (ECL), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA), and Carrière, Frédéric

Subjects
Lipolysis, [SDV]Life Sciences [q-bio], 030303 biophysics, Melting profile, Biochemistry, Intestinal absorption, 03 medical and health sciences, chemistry.chemical_compound, Differential scanning calorimetry, TRACER, Food science, Triolein, Polymorphism, Molecular Biology, Triglycerides, 030304 developmental biology, 0303 health sciences, Molecular Structure, Chemistry, Organic Chemistry, fungi, Temperature, Cell Biology, Lipid, Dietary Fats, X-ray diffraction, [SDV] Life Sciences [q-bio], [SDV.AEN] Life Sciences [q-bio]/Food and Nutrition, Tripalmitin, Anhydrous, Digestion, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition
Abstract

International audience; Dietary fats are present in the diet under different types of structures, such as spread vs emulsions (notably in processed foods and enteral formula), and interest is growing regarding their digestion and intestinal absorption. In clinical trials, there is often a need to add stable isotope-labeled triacylglycerols (TAGs) as tracers to the ingested fat in order to track its intestinal absorption and further metabolic fate. Because most TAG tracers contain saturated fatty acids, they may modify the physicochemical properties of the ingested labeled fat and thereby its digestion. However, the actual impact of tracer addition on fat crystalline properties and lipolysis by digestive lipases still deserves to be explored. In this context, we monitored the thermal and polymorphic behavior of anhydrous milk fat (AMF) enriched in homogeneous TAGs tracers and further compared it with the native AMF using differential scanning calorimetry and power X-ray diffraction. As tracers, we used a mixture of tripalmitin, triolein and tricaprylin at 2 different concentrations (1.5 and 5.7 wt%, which have been used in clinical trials). The addition of TAG tracers modified the AMF melting profile, especially at the highest tested concentration (5.7 wt%). Both AMF and AMF enriched with 1.5 wt% tracers were completely melted around 37 °C, i.e. close to the body temperature, while the AMF enriched with 5.7 wt% tracers remained partially crystallized at this temperature. Similar trends were observed in both bulk and emulsified systems. Moreover, the kinetics of AMF polymorphic transformation was modified in the presence of tracers. While only β’ form was observed in the native AMF, the β-form was clearly detected in the AMF containing 5.7 wt% tracers. We further tested the impact of tracers on the lipolysis of AMF in bulk using a static in vitro model of duodenal digestion. Lipolysis of AMF enriched with 5.7 wt% tracers was delayed compared with that of AMF and AMF enriched with 1.5 wt% tracers. Therefore, low amounts of TAG tracers including tripalmitin do not have a high impact on fat digestion, but one has to be cautious when using higher amounts of these tracers.

Published
2019
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48. INFOGEST static in vitro simulation of gastrointestinal food digestion

Author

Frédéric Carrière, Torsten Bohn, Carla Martins, Adam Macierzanka, Didier Dupont, Werner Weitschies, Sébastien Marze, Ricardo Assunção, Alfonso Clemente, André Brodkorb, Uri Lesmes, Reto Portmann, Cláudia N. Santos, Isabelle Souchon, Milena Corredig, Claire Dufour, David Julian McClements, Claire Bourlieu-Lacanal, Rachel Boutrou, Martin S. J. Wickham, Paula Alvito, Olivia Ménard, Alan R. Mackie, Isidra Recio, Simon Ballance, Catharina Edwards, Gerd E. Vegarud, Mans Minekus, R. Paul Singh, Lotti Egger, Sibel Karakaya, Marie Alminger, Matt Golding, Steven Le Feunteun, Bente Kirkhus, Teagasc Food Research Centre [Fermoy, Ireland], Agroscope, Department of Biology and Biological Engineering, Chalmers University of Technology, Chalmers University of Technology [Gothenburg, Sweden], Instituto Nacional de Saùde Dr Ricardo Jorge [Portugal] (INSA), Nofima AS, Luxembourg Institute of Health, Luxembourg Institute of Healt, Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE), Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université de Montpellier (UM)-Institut National de la Recherche Agronomique (INRA), Science et Technologie du Lait et de l'Oeuf (STLO), Institut National de la Recherche Agronomique (INRA)-AGROCAMPUS OUEST, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), Bioénergétique et Ingénierie des Protéines (BIP ), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Estación Experimental del Zaidín (EEZ), Consejo Superior de Investigaciones Científicas [Madrid] (CSIC), Department of Food Science [Copenhagen] (UCPH FOOD), Faculty of Science [Copenhagen], University of Copenhagen = Københavns Universitet (KU)-University of Copenhagen = Københavns Universitet (KU), Sécurité et Qualité des Produits d'Origine Végétale (SQPOV), Avignon Université (AU)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Quadram Institute Bioscience, Riddet Institute, Department of Food Engineering, Faculty of Engineering, Ege university, Israel Institute of Technology Technion City, Technion City, Faculty of Chemistry, Taras Shevchenko National University of Kyiv, School of Food Science & Nutrition, University of Leeds, Unité de recherche sur les Biopolymères, Interactions Assemblages (BIA), Institut National de la Recherche Agronomique (INRA), Chenoweth Lab, Department of Food Science, University of Massachusetts System (UMASS), Triskelion, Instituto de Biologia Experimental e Tecnológica (IBET), Instituto de Tecnologia Química e Biológica, Génie et Microbiologie des Procédés Alimentaires (GMPA), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Departments of Biological and Agricultural Engineering, and of Food Science and Technology, University of California [San Diego] (UC San Diego), University of California-University of California, Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), Reacta Biotech Limited, Ernst Moritz Arndt, Universität Greifswald - University of Greifswald, Instituto de Investigación en Ciencias de la Alimentación CIAL, CSIC-UAM, CEI UAM+CSIC (CIAL, CSIC-UAM, CEI UAM+CSIC), COST action FA1005 INFOGEST, Teagasc Food Research Centre [Fermoy, County Cork, Ireland], Chalmers University of Technology, Food and Nutrition Department, National Institute of Health Doutor Ricardo Jorge I.P., Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Department of Physiology and Biochemistry of Animal Nutrition, Estacion Experimental del Zaidin, Department of Food Science, University of Guelph, Food Engineering Department, Chimie Et Interdisciplinarité : Synthèse, Analyse, Modélisation (CEISAM), Université de Nantes - Faculté des Sciences et des Techniques, Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS), Department of Biotechnology and Food Engineering, Israel Institute of Technology, Institute of Food Research, Institute of Food Research [Norwich], Université de Minho, Portugal, Laboratoire de Physique des Solides (LPS), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), University of Massachusetts [Amherst] (UMass Amherst), Human Language Technology Interest Group (HULTIG), Universidade de Beira Interior, Instituto de Investigaciòn en Ciencias de la Alimentaciòn (CIAL, CSIC-UAM), Avignon Université (AU)-Institut National de la Recherche Agronomique (INRA), National Institute of Health Doutor Ricardo Jorge, Lisbon and CESAM, University of Aveiro, Estación Experimental del Zaidin, Consejo Superior de Investigaciones Científicas (CSIC), Institut National de la Recherche Agronomique (INRA)-Avignon Université (AU), Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université de Montpellier (UM)-Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (France), French National Institute for Agricultural Research, European Cooperation in Science and Technology (COST), FA1005, Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Norwegian Institute of Food,Fisheries and Aquaculture Research (NOFIMA), University of Copenhagen = Københavns Universitet (UCPH)-University of Copenhagen = Københavns Universitet (UCPH), Quadram Institute Bioscience [Norwich, U.K.] (QIB), Biotechnology and Biological Sciences Research Council (BBSRC), University of California (UC)-University of California (UC), and Ege Üniversitesi

Subjects
Biomimetic materials, [SDV]Life Sciences [q-bio], Oligosaccharides, simulation models, static model, Matrix (chemical analysis), structure de l'aliment, Human health, Eating, 0302 clinical medicine, static in vitro digestion, Biomimetic Materials, [SDV.IDA]Life Sciences [q-bio]/Food engineering, Bile, Food science, Amino Acids, ComputingMilieux_MISCELLANEOUS, 2. Zero hunger, 0303 health sciences, Gastric Juice, Chemistry, Hydrolysis, Fatty Acids, Stomach, Food Ingredients, santé humaine, Hydrogen-Ion Concentration, Intestines, nutrition, Avaliação do Impacte em Saúde, physiological digestion, Digestion, modèle de simulation, Models, Biological, allégations nutritionnelles et de santé, General Biochemistry, Genetics and Molecular Biology, modèle statique, digestion gastrointestinale, 03 medical and health sciences, modèle statique un vitro, simulated digestion, Upper gastrointestinal, Humans, Gastric lipase, Food structure, Saliva, 030304 developmental biology, Enzyme Assays, cinétique de digestion, Mouth, composition de l'aliment, digestion gastrique in vitro, Intestinal digestion, food digestion, Peptide Fragments, Composição dos Alimentos, micronutriment, Food, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, 030217 neurology & neurosurgery
Abstract

WOS: 000462482100001, PubMed ID: 30886367, Developing a mechanistic understanding of the impact of food structure and composition on human health has increasingly involved simulating digestion in the upper gastrointestinal tract. These simulations have used a wide range of different conditions that often have very little physiological relevance, and this impedes the meaningful comparison of results. The standardized protocol presented here is based on an international consensus developed by the COST INFOGEST network. The method is designed to be used with standard laboratory equipment and requires limited experience to encourage a wide range of researchers to adopt it. It is a static digestion method that uses constant ratios of meal to digestive fluids and a constant pH for each step of digestion. This makes the method simple to use but not suitable for simulating digestion kinetics. Using this method, food samples are subjected to sequential oral, gastric and intestinal digestion while parameters such as electrolytes, enzymes, bile, dilution, pH and time of digestion are based on available physiological data. This amended and improved digestion method (INFOGEST 2.0) avoids challenges associated with the original method, such as the inclusion of the oral phase and the use of gastric lipase. The method can be used to assess the endpoints resulting from digestion of foods by analyzing the digestion products (e.g., peptides/amino acids, fatty acids, simple sugars) and evaluating the release of micronutrients from the food matrix. The whole protocol can be completed in similar to 7 d, including similar to 5 d required for the determination of enzyme activities., COST actionEuropean Cooperation in Science and Technology (COST) [FA1005 INFOGEST26]; French National Institute for Agricultural Research (INRA)Institut National de la Recherche Agronomique (INRA), We acknowledge COST action FA1005 INFOGEST26 (http://www.cost-infogest.eu/) for providing funding for travel, meetings and conferences (2011-2015). We acknowledge the French National Institute for Agricultural Research (INRA, http://www.inra.fr) for its continuous support of the INFOGEST network by organizing and co-funding the International Conference on Food Digestion and workgroup meetings. We thank A.G.F. Lopes (Universidade de Lisboa, Portugal) and V.S.N. Mishra (Teagasc Food Research Centre, Moorepark, Ireland) for their help in the final preparation of the videos. We also acknowledge the many other researchers, mostly associated with the above COST action and subsequent events, who have contributed to the discussion of digestion parameters.

Published
2019
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49. Correction: Free fatty acid release from vegetable and bovine milk fat-based infant formulas and human milk during two-phase: In vitro digestion

Author

Lara W. Zeper, Anouk L. Feitsma, Trine Kastrup Dalsgaard, Jeske H.J. Hageman, Arie G. Nieuwenhuizen, Jaap Keijer, and Frédéric Carrière

Subjects
chemistry.chemical_classification, Bovine milk, Chemistry, Humane Voeding & Gezondheid, Fatty acid, General Medicine, In vitro digestion, Phase (matter), Human and Animal Physiology, WIAS, Fysiologie van Mens en Dier, Life Science, Food science, Food Science, VLAG, Human Nutrition & Health
Abstract

The authors regret that the lipid composition of IF1 was reported incorrectly. The percentage of C18:1n-9 should be 42.3%. Since the incorrect value was also used for some calculations, this also affects some of the results: it increases the total amount of fatty acids in the sample, and consequently the percentage of released FFA is lower. The FFA release (as a percentage of initial composition), both of total FFA and C18:1, is similar for both IFs. One small difference between IF1 and IF2 that was seen when using the incorrect value, i.e. a faster early duodenal digestion for IF1, was found to be no longer statistically significant. This has no consequences for the conclusions of the manuscript. Page 2106 should read " The human milk samples showed less release of FFA during the gastric phase compared to IF1 and IF2 (2.0 ± 0.2% vs. 4.3 ± 0.2% and 4.7 ± 0.1% respectively, p < 0.01). Compared to the amount of FFA released after the digestion, during the gastric phase 4% of FFA were released from human milk, about 10% from IF1, and about 11% from IF2. Except for 45 minutes (p = 0.04), i.e. 15 minutes after the start of the duodenal phase, no differences were found in FFA release between the IFs compared to the human milk samples during this phase. The total release of FAs at the end of the digestion, as percentage of initial composition, was found to be similar for the different samples (43.9 ± 2.0%, 42.2 ± 1.4%, and 52.3 ± 4.5% for IF1, IF2 and human milk respectively, p = 0.14)". The correspondingly updated Fig. 2B, Fig. 5K, Table 2 and Table 3 are as presented below.(Table Persented).

Published
2019
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50. Free fatty acid release from vegetable and bovine milk fat-based infant formulas and human milk during two-phase in vitro digestion

Author

Frédéric Carrière, Arie G. Nieuwenhuizen, Jeske H.J. Hageman, Jaap Keijer, Anouk L. Feitsma, Trine Kastrup Dalsgaard, Lara W. Zeper, Wageningen University and Research [Wageningen] (WUR), Department of Food Science, Bioénergétique et Ingénierie des Protéines (BIP ), and Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Infant Formula/chemistry, [SDV]Life Sciences [q-bio], Regulation of gastric function, Absorption (skin), Fatty Acids, Nonesterified, Models, Biological, 03 medical and health sciences, In vivo, Vegetables/chemistry, Vegetables, Milk/chemistry, Lipolysis, Life Science, Animals, Humans, Food science, VLAG, Human Nutrition & Health, 2. Zero hunger, chemistry.chemical_classification, 030109 nutrition & dietetics, Milk, Human, Chemistry, Humane Voeding & Gezondheid, Fatty acid, Infant, Milk, Human/chemistry, food and beverages, General Medicine, Infant Formula, Fatty Acids, Nonesterified/chemistry, Renal disorders Radboud Institute for Molecular Life Sciences [Radboudumc 11], 030104 developmental biology, Milk, Infant formula, Human and Animal Physiology, WIAS, Fysiologie van Mens en Dier, Composition (visual arts), Cattle, Digestion, Female, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Food Science
Abstract

Item does not contain fulltext BACKGROUND: Bovine milk fat is increasingly used in infant formula (IF). The triacylglycerol (TAG) structure of bovine milk fat might be beneficial for digestion and absorption. We investigated the release of fatty acids (FAs) of IF containing different fat blends and compared this to human milk. METHODS: Fresh human milk was sampled and two IFs were produced; one containing 100% vegetable fat (IF1) and one with 67% bovine milk fat and 33% vegetable fat (IF2). Using a static in vitro infant digestion model, consisting of a gastric and duodenal phase, the time dependent release of individual free fatty acids (FFA) was studied and analysed using GC-MS, and residual TAG levels were determined by GC-FID. RESULTS: Human milk and the IFs showed comparable total FA release. In the gastric phase, 4-11% of lipolysis occurred, and mainly short (SCFA)- and medium chain fatty acids (MCFA) were released. In the duodenal phase, lipolysis proceeded with release of C4:0 but was marked by a fast release of long-chain fatty acids (LCFA). The digestion of the IFs resulted in different FFA profiles during and at the end of digestion. IF2 gave more release of C4:0-C11:0, which reflects the FA composition of bovine milk. CONCLUSION: The addition of bovine milk fat to IF resulted in a total FA release comparable to an IF with only vegetable fat and human milk. However, it did lead to a different time-dependent release of individual FAs, which might result in differences in absorption and other health effects in vivo.

Published
2019
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