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Lupins, and other legumes, have attained international interest due to their reported remarkable health benefits. Currently, the seed coats are discarded as waste or animal feed. The research presented here summarizes the potential for incorporating the seed coats into 'whole grain' foods. We aimed to identify metabolites found in the seed coats of nine commercial Australian cultivars of lupin (Lupinus angustifolius and L. albus species), and to evaluate and compare their functional, nutritional, antioxidant, and antidiabetic properties, along with in silico exploration of mechanisms of action for selected identified secondary metabolites. The seed coats were found to contain 79 to 90% dietary fibers and substantial quantity of essential macrometals. LC-QTOF MS-based, untargeted bioactive metabolite profiling explored a total of 673 chemical entities, and identified 63 bioactive secondary metabolites including: biophenols, unsaturated fatty acids, triterpenoids, alkaloids, and dietary prebiotics (insoluble fibers). The seed coats from these nine cultivars show substantial antioxidant activity. The cultivars of L. angustifolius inhibit α-amylase and α-glucosidase significantly in vitro. Moreover, in silico docking and dynamic simulation along with ADME/T analysis suggest that quercetin 3-methyl ether and 8-C-methylquercetin 3-methyl ether as molecules, novel in lupin seed coats, are responsible for the α-amylase and α-glucosidase inhibition. The findings indicated that lupin seed coats might be beneficial food components, rather than be discarded as 'mill waste'., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Surface tension (γ eq ) of the seed extracts of four lupine cultivars showed values in the range 44.9-46.4 mN/m. The surface compression elasticity (E') of the adsorbed layers and foaming capacity (FC) also showed similar values (E' ∼ 30 mN/m, FC ∼ 100%). The effect of defatting prior to extraction at pH 8.5 depends on the solvent employed - hexane and dichloromethane improved the subsequent protein extraction yield, while ethanol reduced it. The effect of defatting on surface tension could be positive (for hexane and ethanol) or negative (for dichloromethane). Generally, defatting improved the surface compression rheological and foaming parameters. On the other hand, fractionation of the extracts obtained at pH 8.5 from hexane-defatted seeds did not improve significantly the surface activity parameters. Some improvement with respect to the unfractionated extracts was observed only for the extracts of undefatted seeds. γ eq , E', E" and FC isotherms confirm the surfactant-like behavior of the lupine seed extracts., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In this work we examined the contents of 14 polyphenols in white lupin ( Lupinus albus L.) samples cultivated in Southern Italy by the optimisation and validation of a LC-HRMS method. The validation of the LC-HRMS method showed linearity results r 2 > 0.989 and recovery values between 71 and 119% for a very wide range of concentrations. Ellagic acid was the most abundant polyphenol, with mean concentrations of 16271.86 ± 19798.53 μg/Kg, followed by apigenin (2749.51 ± 889.95 μg/Kg). A significant variability in ellagic acid contents was found between the areas of cultivation examined ( p  < 0.05). As far as we know, this work provides the first data on the polyphenols contents of white lupins cultivated in Italy. The comparison with other study confirms the role of the cultivation area for the determination of the polyphenol's contents. The study also confirms white lupins as a promising source of antioxidant and anti-inflammatory substances in a balanced diet.

Lupin holds an important place among the legumes and the utilization of lupin as a dietary protein source is an excellent environmentally friendly alternative to animal-based products for human nutrition. In the present study, nutritional, thermal, rheological and functional properties of nine Australian lupin cultivars have been assayed in order to find the most valuable one, both nutritiously and industrially. The set comprised six Lupinus angustifolius L. viz., Barlock, Gunyadi, Jenabillup, Jindalee, Jurien, Mandelup and three Lupinus albus L. viz., Luxor, Rosetta, WK388 cultivars. The tests included analysis of color, macronutrient and micronutrient composition, pasting, textural and thermal properties, electrophoretic profile of protein isolates, swelling power, water and oil absorption capacity, emulsifying capacity, emulsion stability, creaming stability, foaming capacity and stability of the cultivars' dehulled seed flours. The results indicated substantial variation in macro and micro-nutritional value as well as satisfactory swelling ability, solubility, surface hydrophobicity, foaming ability, emulsifying capacity and gelation property of lupin flours. Superior nutritional, thermal, rheological and functional potential was demonstrated by the L. albus cultivars compared to the L. angustifolius cultivars with the exception of Mandelup., (© 2021. The Author(s).)

There is a renewed interest on the reliance of food-based bioactive compounds as sources of nutritive factors and health-beneficial chemical compounds. Among these food components, several proteins from foods have been shown to promote health and wellness as seen in proteins such as α/γ-conglutins from the seeds of Lupinus species (Lupin) , a genus of leguminous plant that are widely used in traditional medicine for treating chronic diseases. Lupin-derived peptides (LDPs) are increasingly being explored and they have been shown to possess multifunctional health improving properties. This paper discusses the intestinal transport, bioavailability and biological activities of LDPs, focusing on molecular mechanisms of action as reported in in vitro, cell culture, animal and human studies. The potentials of several LDPs to demonstrate multitarget mechanism of regulation of glucose and lipid metabolism, chemo- and osteoprotective properties, and antioxidant and anti-inflammatory activities position LDPs as good candidates for nutraceutical development for the prevention and management of medical conditions whose etiology are multifactorial.

Fermented soy sauces are used as food seasonings in Eastern countries and all over the world. Depending on their cultural origins, their production differs in parameters such as wheat addition, temperature, and salt concentration. The fermentation of lupine seeds presents an alternative to the use of soybeans; however, the microbiota and influencing factors are currently unknown. In this study, we analyse the microbiota of lupine Moromi (mash) fermentations for a period of six months and determine the influence of different salt concentrations on the microbiota dynamics and the volatile compound composition. Cultured microorganisms were identified by protein profiling using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS), and 16S rRNA gene amplicon sequencing provided an overview of the microbiota including non-cultured bacteria. The volatile compounds were determined by gas chromatography-mass spectrometry (GC-MS). At all salt concentrations, we found that Tetragenococcus halophilus (up to 1.4 × 10 9 colony forming units (CFU)/mL on day 21) and Chromohalobacter japonicus (1.9 × 10 9  CFU/mL, day 28) were the dominating bacteria during Moromi fermentation. Debaryomyces hansenii (3.6 × 10 8  CFU/mL, day 42) and Candida guilliermondii (2.2 × 10 8  CFU/mL, day 2) were found to be the most prevalent yeast species. Interestingly, Zygosaccharomyces rouxii and other yeasts described as typical for soy Moromi were not found. With increasing salinity, we found lower diversity in the microbiota, the prevalence-gain of typical species was delayed, and ratios differed depending on their halo- or acid tolerance. GC-MS analysis revealed aroma-active compounds, such as pyrazines, acids, and some furanones, which were mostly different from the aroma compounds found in soy sauce. The absence of wheat may have caused a change in yeast microbiota, and the use of lupine seeds may have led to the differing aromatic composition. Salt reduction resulted in a more complex microbiome, higher cell counts, and did not show any spoiling organisms. With these findings, we show that seasoning sauce that uses lupine seeds as the sole substrate is a suitable gluten-free, soy-free and salt reduced alternative to common soy sauces with a unique flavour., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Lupin seeds can represent a valuable source of phenolics and other antioxidant compounds. In this work, a comprehensive analysis of the phytochemical profile was performed on seeds from three Lupinus species, including one cultivar ( Lupinus albus ) and two wild accessions ( Lupinus cossentinii and Lupinus luteus ), collected from the northern region of Tunisia. Untargeted metabolomic profiling allowed to identify 249 compounds, with a great abundance of phenolics and alkaloids. In this regard, the species L. cossentinii showed the highest phenolic content, being 6.54 mg/g DW, followed by L. luteus (1.60 mg/g DW) and L. albus (1.14 mg/g DW). The in vitro antioxidant capacity measured by the ABTS assay on seed extracts ranged from 4.67 to 17.58 mg trolox equivalents (TE)/g, recording the highest values for L. albus and the lowest for L. luteus . The DPPH radical scavenging activity ranged from 0.39 to 3.50 mg TE/g. FRAP values varied between 4.11 and 5.75 mg TE/g. CUPRAC values for lupin seeds ranged from 7.20 to 8.95 mg TE/g, recording the highest for L. cossentinii . The results of phosphomolybdenum assay and metal chelation showed similarity between the three species of Lupinus . The acetylcholinesterase (AChE) inhibition activity was detected in each methanolic extract analyzed with similar results. Regarding the butyrylcholinesterase (BChE) enzyme, it was weakly inhibited by the Lupinus extracts; in particular, the highest activity values were recorded for L. albus (1.74 mg GALAE/g). Overall, our results showed that L. cossentinii was the most abundant source of polyphenols, consisting mainly in tyrosol equivalents (5.82 mg/g DW). Finally, significant correlations were outlined between the phenolic compounds and the in vitro biological activity measured, particularly when considering flavones, phenolic acids and lower-molecular-weight phenolics.

Plant derived fermented beverages have recently gained consumers' interest, particularly due to their intrinsic functional properties and presence of beneficial microorganisms. Three variants containing 5%, 10%, and 15% ( w / w ) of sweet blue lupin ( Lupinus angustifolius L. cv. "Boregine") seeds were inoculated with kefir grains and incubated at 25 °C for 24 h. After processing, beverages were stored in refrigerated conditions (6 °C) for 21 days. Changes in microbial population, pH, bioactive compounds (polyphenolics, flavonoids, ascorbic acid), reducing sugars, and free amino acids were estimated. Additionally, viscosity, firmness, color, and free radicals scavenging properties were determined. Results showed that lactic acid bacteria as well as yeast were capable of growing well in the lupin matrix without any supplementation. During the process of refrigeration, the viability of the microorganisms was over the recommended minimum level for kefir products. Hydrolysis of polysaccharides as well as increase of free amino acids was observed. As a result of fermentation, the beverages showed excellent DPPH, ABTS +· , · OH, and O 2 - radicals scavenging activities with a potential when considering diseases associated with oxidative stress. This beverages could be used as a new, non-dairy vehicle for beneficial microflora consumption, especially by vegans and lactose-intolerant consumers.

Ceramides (Cer) and cerebrosides are important sphingolipids (SL) involved in many biological processes. Herein, the SL content of yellow lupin seeds (Lupinus luteus) was determined by liquid-liquid extraction, mild alkaline hydrolysis (1 h at 37 °C) and reversed-phase liquid chromatography with negative electrospray ionization coupled to either an orbital-trap (Fourier-transformed, FT) or linear ion-trap (LIT) mass spectrometry (RPLC-ESI/MS). The chemical identity of SL including the sugar residues and sphingoid backbone (SB) was inferred by collision-induced dissociation multiple-stage mass spectrometry (MS n , n = 2,3). Up to 52 Cer and 47 cerebrosides were successfully recognized and quantified in sample extracts of L. luteus seeds also counting isobaric species. As reported for other vegetables, a hydroxylated SB was observed whereby the N-acyl chains showed a high occurrence of very-long-chain moieties; phytosphingosine and 4-hydroxy-8-sphingenine were the predominant SB paired mainly with oleic acid (C18:1), hydroxylated behenic acid (C22:0;1) and hydroxylated lignoceric acid (C24:0;1)., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

1,2-Diacyl- sn -glycero-3-phospho- N -acyl-ethanolamines (NAPE) are low abundance phospholipids but important constituents of intracellular membranes of plant tissues, responsible for generating bioactive N -acylethanolamine (NAE), which participates in several physiological processes such as regulation of seed germination and protection against pathogenic attacks. From an analytical point of view, the critical aspect of these bioactive lipids lies in the determination of fatty acyl chains located in sn-1/sn-2 position on the glycerol backbone ( O -linked), along with the amide-bound ( N -linked) fatty acyl chain. Here, the identity and occurrence of NAPE in lipid extracts of lupin seeds ( Lupinus luteus L.) was assessed by electrospray ionization in negative ion mode upon reversed-phase liquid chromatography (RPLC-ESI) coupled to mass spectrometry (MS) either at high- (i.e., Orbitrap FTMS) or low- (linear ion trap, LIT) resolution/accuracy. Collisional induced dissociation (CID)-tandem MS and MS 3 acquisitions of chemically prepared NAPE allowed to unequivocally recognize the N -linked fatty acyl chain and to establish the diagnostic product ions that were successfully applied to identify NAPE in lipid extracts of yellow lupin seeds. The most abundant NAPE species were those containing N -acyl groups C18:1, C18:2; a minor prevalence was found for C16:0, C18:0, and C18:3, and almost the same acyl chains O -linked on the glycerol backbone in several sn-1/sn-2 combinations were observed. The positional isomers of NAPE species were identified as deprotonated molecules ([M-H] - ) at m / z 978.7541 (three isomers 52:3 ), m / z 980.7694 (two isomers 52:2 ), m / z 1002.7535 (four isomers 54:5 ), m / z 1004.7686 (two isomers 54:4 ), m / z 1006.7837 (two isomers 54:3 ), and m / z 1008.8026 (single isomer 54:2 ). The total amount of NAPE in lupin seeds ranged in the interval of 2.00 ± 0.13 mg/g dw, in agreement with other edible legumes. We anticipate our approach to be a robust assessment method potentially applicable to biological extracts containing NAPE species and can provide comprehensive profiles and contents.

Lupin seeds are rich in proteins and other essential ingredients that can help to improve human health. The protein contents in both whole and split seeds of two lupin cultivars (Mandleup and PBA Jurien) were used to produce the lupin milk using the cheesecloth and centrifuge method. Proteins were extracted from the lupin milk using thiourea/urea solubilization. The proteins were separated by a two-dimensional polyacrylamide gel electrophoresis and then identified with mass spectrometry. A total of 230 protein spots were identified, 60 of which showed differential abundances. The cheesecloth separation showed protein extractability much better than that of the centrifuge method for both the cultivars. The results from this study could offer guidance for future comparative analysis and identification of lupin milk protein and provide effective separation technique to determine specific proteins in the cheese-making process.

Background: The presence of quinolizidine alkaloids (QAs) in the species Lupinus mutabilis Sweet limits the expansion of its consumption and use, despite its high protein content. The objective of this research was therefore to determine the effect of two thermal treatments, aqueous (ATT) and saline (STT), on the QAs and total protein content, as well as on the texture (fracturability and hardness), visual perception attributes - hue (H*), luminosity (L*) and chromatism (C*) - and grain size in three lupin varieties (INIAP-450, INIAP-451, and Criollo). The water consumption required by each treatment was also measured., Results: The debittering process with ATT helped to concentrate the total nitrogen by 560 g kg -1 and decreased the grain hardness to 2037 gf (grams of force) in the Criollo variety, while the chromatic parameters H* and C* increased in the three varieties. The STT treatment was more efficient than the ATT treatment in terms of the time required and the volume of water used to reduce the QAs to safe levels for consumption (2.5-3.5 g kg -1 ). The size of the grain increased to four times its original size; the luminosity L* decreased during cooking to a value of 41.49 in the Criollo variety and then increased to 57.42 during grain washing., Conclusions: The STT treatment is advisable for lupin debittering, although the extent of the effect was dependent on the variety. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Metabolic changes occurring in white lupine grain were investigated in response to Plant Growth Promoting Rhizobacteria (PGPR) root inoculation under field condition. We precisely targeted lipids and phenolics changes occurring in white lupine grain in response to Pseudomonas brenneri LJ215 and/or Paenibacillus glycanilyticus LJ121 inoculation. Lipids and phenolic composition were analyzed using an Ultra High-Performance Liquid Chromatography/Tandem Mass Spectrometry Methods. As compared to grain of un-inoculated control plant, Paenibacillus glycaniliticus inoculation highly increased the total lipids content (from 232.55 in seeds of un-inoculated control plant to 944.95 mg/kg) and the relative percentage of several fatty acid such as oleic acid (+20.95%) and linoleic acid (+14.28%) and decreased the relative percentage of glycerophospholipids (- 13.11%), sterol (- 0.2% and - 0.34% for stigmasterol and campesterol, respectively) and prenol (- 17.45%) class. Paenibacillus glycaniliticus inoculation did not affect total phenolic content, while it modulated content of individual phenolic compounds and induced the accumulation of "new" phenolics compounds such as kaempferol. Paenibacillus glycanilyticus LJ121 can be a useful bio-fertilizer to enhance nutritional quality of white lupine grain.

Lupinus angustifolius L. is a legume culture known as a source of valuable feed protein and the N 2 -fixator for improving soil fertility. However, its low ecological resistance does not allow for a stable yield of the crop. Earlier, we have shown that steroid phytohormone 24-epibrassinolide (EBR) increases the tolerance of lupine to chlorine ions by activating the protective proteins in ripening seeds (such as proteinase inhibitors that prevent protein breakdown) and lectins. Here we investigated the effect of EBR on the functional status of the N 2 -fixing system in root nodules, protein synthesis in ripening seeds and the resistance of lupine plants to various pathogens. It was found that EBR enhanced the nodulation process, N 2 -fixing activity of nitrogenase and the accumulation of poly-β-hydroxybutirate in the bacteroides, increased the leghemoglobin content in the nodules as well as the metabolic activity of bacteroides. According to data on the inclusion of 14 C-leucine in maturing seed proteins, EBR increased the accumulation of protein in them against the background of a short-term decrease in protein synthesis and its subsequent regeneration to the control level. Gradual inhibition of protein synthesis, characteristic of other legumes, was observed in control variants of lupine. EBR increased lupine resistance to phytopathogenic fungi of Colletotrichum genus and insects of Noctuidae and Scarabaeidae families. We concluded that a more complete implementation of the potential productivity and sustainability of lupine under the action of EBR was achieved due to the anabolic/anti-catabolic effect on the N 2 fixation system in root nodules, as well as on protein synthesis in ripening seeds., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Seed extracts of Nymphia alba Linn. and Lupinus polyphyllus Lindl. were analyzed for fatty acid composition, functional group analysis and antioxidant activity. The petroleum ether extract of seeds were found dominant in unsaturated fatty acids with oleic acid (39.9%) and linoleic acid (29.6%) in L. polyphyllus and linoleic (37.5%) and oleic acid (10.9%) in N. alba. All the defatted seed extracts of N. alba and L. polyphyllus found to have powerful DPPH, ABTS, H 2 O 2 and NBT antioxidant radical scavenging activity with reference to butylated hydroxy toluene (BHT). The defatted seed extracts were further analyzed with functional group analysis through FTIR found to contain numerous functional groups which may be responsible for their antioxidant activity.

In this study, some physico-chemical properties, amino acids, fatty acids, sugars and mineral contents of sorghum, millet and lupin seeds. Sorghum (red, white and yellow) and millet seeds were purchased from market in Saudi Arabia (Riyadh). Lupin seeds were provided from in Turkey (Konya). Protein contents of seed samples ranged from 8.6% (yellow sorghum) to 37.7% (lutop) (p < 0.05). The extractable phenolics contents for gallic acid equivalent (GAE) of grains ranged between 1.43 mgGAE/g (white sorghum) to 8.23 mgGAE/g (red sorghum), and hydrolysable phenolics contents for GAE of grains varied between 1.48 mgGAE/g (white sorghum) to 26.10 mgGAE/g (red sorghum (p < 0.05). Total phenol contents of seeds were found between 2769 mg GAE/g (bablon) to 6087 mgGAE/g (yellow sorghum) (p < 0.05). Amino acid contents of millet changed between 0.02% (ornithine) and 2.07% (glutamic acid), while amino acid contents of yellow sorghum range from 0.02% (hydroxyproline) to 1.71% (glutamic acid), amino acid values of white sorghum changed between 0.02% (hydroxyproline) and 2.21% (glutamic acid), amino acid values of lutop seed changed between 0.02% (ornithine) and 6.77% (glutamic acid) (p < 0.05).While the oleic acid contents change between 25.27% (white sorghum) and 53.50% (Bablone), linoleic acid contents ranged from 14.60% (Bablone) to 42.67% (Millet) (p < 0.05). However, the amount of potassium in the seeds varied between 1831.34 mg/kg (while sorghum) and 11895.8 mg/kg (Lutop). Generally, protein, oleic acid, amino acid and mineral contents of lupin varieties were higher as compared to those of millet phenol, anthocyanin and sorghum seeds.

The quality and amount of yellow lupine yield depend on water availability. Water scarcity negatively affects germination, flowering, and pod formation, and thus introduction of an artificial irrigation system is needed. The aim of this study was to evaluate the influence of irrigation on the quality of yellow lupine seeds. Raining was applied with a semi-solid device with sprinklers during periods of greatest water demand. It was shown that watered plants produced seeds of lesser quality, having smaller size and weight. To find out why seeds of irrigated plants were of poor quality, interdisciplinary research at the cellular level was carried out. DNA cytophotometry evidenced the presence of nuclei with lower polyploidy in the apical zone of mature seeds. This may lead to formation of smaller cells and reduce depositing of storage materials. The electrophoretic and Fourier transform infrared spectroscopic analyses revealed differences in protein and cuticular wax profiles, while scanning electron microscopy and energy dispersive spectroscopy revealed, among various chemical elements, decreased calcium content in one of seed zones (near plumule). Seeds from irrigated plants showed slightly higher germination dynamics but growth rate of seedlings was slightly lower. The studies showed that irrigation of lupine affected seed features and their chemical composition, an ability to germination and seedlings growth.

1 . This study investigated the effect of different levels of seeds of lupin species ( Lupinus angustifolius, Lupinus luteus and Lupinus albus ) on broiler growth performance, nutrient digestibility and apparent metabolisable energy of poultry diets. 2 . A control diet containing corn-soybean meal and test diets containing 50, 100, 150, 200 and 250 g/kg of three lupin species were fed to broiler chickens during a 35-day study period. There were 10 replicate cages of 8 broiler chickens per treatment. 3 . Growth performance of the birds fed with yellow and white lupin diets was found to be similar to that of the birds in the control group. The use of narrow-leaved lupin seeds in the diet quadratically decreased the body weight gain of birds and increased feed conversion ratio during all the experimental periods (0-14, 15-35 and overall 0-35 days of age). 4 . A significant interaction between lupin species and their inclusion levels for apparent metabolisable energy and apparent ileal digestibility of starch was observed, showing a negative effect of increasing levels of narrow-leaved lupin. Pearson correlation coefficients demonstrated that the presence of raffinose and non-starch polysaccharides is a key factor affecting the nutritive value of lupin, 5 . Yellow and white lupins could be utilised as an alternative ingredient for soybean meal in the diet of broiler chickens without any adverse effects on growth performance. The narrow-leaved counterpart diet reduced broiler performance, and hence did not provide a satisfactory outcome.

Phenolic compounds are secondary plant metabolites whose beneficial health effects make them of intense interest to researchers. The aim of the study presented here was to evaluate the change in the phenolic compound profile of lupin seed in in vitro digestion. The most abundant phenolic compounds in undigested lupin seeds were mostly apigenin derivatives. The in vitro digestion of lupin seeds resulted in qualitatively altered phenolic compound profiles. Approximately 80% of phenolic compounds were released from lupin seeds during the in vitro digestion, which simulated gastric processes. Continued digestion, imitating the intestinal phase, additionally increased the bioaccessibility of lupin seed polyphenols by about 10%. The in vitro gastrointestinal model was also used to elucidate how the content of native phenolic compounds affects the digestion susceptibility of lupin seed proteins. An inverse correlation between protein digestibility and phenolic compound content, was also demonstrated., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Seeds of the legume lupin (Lupinus spp.) are becoming increasingly important as human food. The seed coat, at ~25% of the whole seed of Lupinus angustifolius (Australian sweet lupin, ASL), is the main by-product of lupin kernel flour production. The primary market for lupin seed coat is low value feed with very limited use in foods. In this study, seed coats of six ASL commercial varieties from two growing sites were sampled for identification and quantification of polyphenols using a high-performance liquid chromatography (HPLC) with diode array detector (DAD) and coupled with a triple quadrupole mass spectrometer which equipped with electrospray ionization source (ESI-MS/MS). Three flavones (apigenin-7-O-β-apiofuranosyl-6,8-di-C-β-glucopyranoside, vicenin 2, and apigenin-7-O-β-glucopyranoside), one isoflavone (genistein) and one dihydroflavonol derivative (aromadendrin-6-C-β-d-glucopyranosyl-7-O-[β-D-apiofuranosyl-(1 → 2)]-O-β-D-glucopyranoside), and several hydroxybenzoic and hydroxycinnamic acid derivatives were identified. Considerable variations in levels of individual polyphenols were found but apigenin-7-O-β-apiofuranosyl-6,8-di-C-β-glucopyranoside was the predominant polyphenol in all samples accounting for 73.08-82.89% of the total free polyphenols. These results suggest that ASL seed coat could be valuable dietary source of polyphenols., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Lupin seed proteins have been reported to exhibit hypoglycaemic effects in animals and humans following oral administration, however little is known about its mechanism of action. This study investigated the signalling pathway(s) responsible for the insulinotropic effect of the hydrolysate obtained from lupin (Lupinus angustifolius L.) seed extracts utilizing BRIN-BD11 β-cells. The extract was treated with digestive enzymes to give a hydrolysate rich in biomolecules ≤7 kDa. Cells exhibited hydrolysate induced dose-dependent stimulation of insulin secretion and enhanced intracellular Ca 2+ and glucose metabolism. The stimulatory effect of the hydrolysate was potentiated by depolarizing concentrations of KCl and was blocked by inhibitors of the ATP sensitive K + channel, Gα q protein, phospholipase C (PLC) and protein kinase C (PKC). These findings reveal a novel mechanism for lupin hydrolysate stimulated insulin secretion via Gα q mediated signal transduction (Gα q /PLC/PKC) in the β-cells. Thus, lupin hydrolysates may have potential for nutraceutical treatment in type 2 diabetes., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The efficient production of saponin-rich extracts is of increasing interest due to the bioactive properties that have being demonstrated for these compounds. However, saponins have a poor bioavailability. In this respect, the knowledge about the bioaccessibility of saponins as a first step before bioavailability has been scarcely explored. In this study, the production of ultrasound-assisted extracts of saponins from edible seeds (quinoa, soybean, red lentil, fenugreek and lupin) was carried out with ethanol, ethanol:water or water. Extraction yield, total saponin (TSC), fat and total phenolics content (TPC) were determined. Then, the bioaccessibility of saponins after the in vitro gastrointestinal digestion of the extracts was determined and the effect of TPC and fat in the extracts on bioaccessibility was evaluated. The highest saponin-rich extracts were obtained by ethanol, being fenugreek and red lentil the richest extracts (12% and 10%, respectively). Saponins from ethanol:water extracts displayed variable bioaccessibility (from 13% for fenugreek to 83% for lentil), but a bioaccessibility closer to 100% was reached for all ethanol extracts. Correlation studies showed that TPC of the extracts negatively affected the bioaccessibility of saponins, whereas fat of the extracts enhanced this parameter. As summary, ultrasound-assisted extraction is shown as an efficient method for obtaining saponin-rich extracts from edible seeds, being ethanol the most advantageous solvent due to the richness of saponins and the successful bioaccessibility from these extracts, likely caused by the co-extracted fat with ethanol. Regardless of the extracts, phenolic compounds or fat may hinder or enhance the bioaccessibility of saponins, respectively. Additionally, an adequate balance between saponins to lipids has shown to be relevant on such an effect., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

This work aimed to extend our current knowledge by obtaining novel information regarding the effect of the dietary inclusion of dehulled white lupine seeds (DWL) on the physical characteristics and sensory qualities of rabbit meat, as well as growth and carcass traits. The DWL diet was compared with a soybean meal-based diet. Feeding the DWL diet to rabbits led to lower Warner-Bratzler shear force values measured in grilled meat samples (Longissimus lumborum muscle) across fibers and this finding was confirmed in the sensory evaluation of texture in terms of higher tenderness and fibrosity of meat samples. No differences were detected by panelists in terms of intensity of aroma, the presence of a flavor typical for cooked meat, or the presence of a flavor typical for rabbit meat. Additionally, saturation and thrombogenic indexes from hind leg meat of rabbits fed the DWL diet were reduced. There was a better feed conversion ratio in rabbits fed the DWL diet., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

In order to promote the use of lupin in pig nutrition, in this research the nutritional characteristics (i.e. dietary fibre, alkaloid and fatty acid profile) and the in vitro gas production of 12 lupin varieties grown in the Mediterranean basin and belonging to three lupin species (Lupinus albus, Lupinus angustifolius and Lupinus luteus) were assessed. Four varieties of L. albus (Asfer, Lublanc, Lutteur and Multitalia) were grown in South Campania. Three varieties of L. luteus (Dukat, Mister and Taper), three of L. angustifolius (Jindalee, Sonet and Wonga) and two of L. albus (Rosetta and Luxor) were grown in Eastern Sicily. Lupinus albus varieties showed interesting nutritional and dietetic characteristics (i.e. high protein and low fibre content); the lipid fraction, rather elevated, is well represented by monounsaturated fatty acids (544 g/kg), whereas saturated fatty acids (SFAs) are less represented (167 g/kg) and the n-3/n-6 ratio (0.510) is the most favourable. Lupinus luteus varieties presented the most remarkable dietetic aspects, in terms of polyunsaturated fatty acid (PUFA) content (569 g/kg), n-6 PUFA series (490 g/kg), UFA/SFA (5.24) and PUFA/SFA (3.56) ratios and atherogenic (0.059) and thrombogenic (0.100) indices and very low alkaloid content (1.07 mg per 100 g). Lupinus angustifolius varieties showed the least interesting nutritional and dietetic characteristics: low protein and fat content, high fibre level, high SFA amount (248 g/kg) and the lowest favourable nutritional indices (IA: 0.164 and IT: 0.334). Regarding the fermentation process, in L. albus, the tendency to increase the rate of gas production during the early stages of fermentation suggests that the high presence of alkaloids did not affect the in vitro degradability, production of short-chain fatty acids and fermentation process, probably due to their concentration and/or water solubility. Lupinus angustifolius and L. luteus showed intermediate and slightly worse in vitro fermentation patterns respectively. From a nutritional and dietetic point of view, lupin may represent an interesting alternative to soya bean in pig feeding., (Journal of Animal Physiology and Animal Nutrition © 2017 Blackwell Verlag GmbH.)

This study investigated the effect of germination and fermentation on the composition of carbohydrates in Australian sweet lupin. Specifically, the amount of sugars (sucrose, fructose, and glucose), starch, oligosaccharides (verbascose, stachyose, and raffinose), and dietary fiber were measured in germinated lupin seeds and fermented lupin flour, and compared with those in soy. High performance liquid chromatography coupled with refractive index was employed for quantitation of sugars, starch, and oligosaccharides, and gas chromatography coupled with a flame ionization detector was used for quantitation of simple sugars in total, and soluble, and insoluble dietary fiber. The enzyme activities of α-amylase and α-glucosidase were compared before and after germination or fermentation. The α-amylase activity in germinated lupin increased to  ∼17 nmol/mL/min/0.1 g and in germinated soy∼32; in fermented lupin, the activity increased to ∼52, while in fermented soy it decreased to ∼20. In general, germination or fermentation decreased the oligosaccharide content, and increased the total sugar in samples (p < 0.05). Total oligosaccharides in lupin after uncontrolled germination were reduced by 98% to 6 mg/g, and after controlled germination reduced by 44% to 86 mg/g. Fermentation with yogurt culture lowered the content of total oligosaccharides due to 94% decrease in stachyose. Total oligosaccharides in soy flour prior to fermentation were 180 mg/g and significantly decreased to ∼124 mg/g in fermented soy. Germination did not affect the starch content. There was no significant change in the amounts of total, soluble, and insoluble dietary fiber after germination or fermentation of lupin except for galactose, which was significantly reduced in germinated lupin seeds. Soluble dietary fiber in germinated soy significantly increased. Germination and fermentation are simple and effective techniques to reduce the oligosaccharides while maintaining the composition of dietary fibers.

A simple, selective and accurate reverse phase HPLC method was developed for detection and quantitation of γ-conglutin from lupin seed extract. A linear gradient of water and acetonitrile containing trifluoroacetic acid (TFA) on a reverse phase column (Agilent Zorbax 300SB C-18), with a flow rate of 0.8ml/min was able to produce a sharp and symmetric peak of γ-conglutin with a retention time at 29.16min. The identity of γ-conglutin in the peak was confirmed by mass spectrometry (MS/MS identification) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The data obtained from MS/MS analysis was matched against the specified database to obtain the exact match for the protein of interest. The proposed method was validated in terms of specificity, linearity, sensitivity, precision, recovery and accuracy. The analytical parameters revealed that the validated method was capable of selectively performing a good chromatographic separation of γ-conglutin from the lupin seed extract with no interference of the matrix. The detection and quantitation limit of γ-conglutin were found to be 2.68μg/ml and 8.12μg/ml respectively. The accuracy (precision and recovery) analysis of the method was conducted under repeatable conditions on different days. Intra-day and inter-day precision values less than 0.5% and recovery greater than 97% indicated high precision and accuracy of the method for analysis of γ-conglutin. The method validation findings were reproducible and can be successfully applied for routine analysis of γ-conglutin from lupin seed extract., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Lupinus mutabilis has protein (32.0-52.6 g/100 g dry weight) and lipid (13.0-24.6 g/100 g dry weight) contents similar to soya bean (Glycine max). The Ω3, Ω6, and Ω9 contents are 1.9-3.0, 26.5-39.6, and 41.2-56.2 g/100 g lipid, respectively. Lupins can be used to fortify the protein content of pasta, bread, biscuits, salads, hamburgers, sausages, and can substitute milk and soya bean. Specific lupin protein concentrates or isolates display protein solubility (>90%), water-absorption capacity (4.5 g/g dry weight), oil-absorption capacity (3.98 g/g), emulsifying capacity (2000 mL of oil/g), emulsifying stability (100%, 60 hours), foaming capacity (2083%), foaming stability (78.8%, 36 hours), and least gelation concentration (6%), which are of industrial interest. Lupins contain bitter alkaloids. Preliminary studies on their toxicity suggest as lethal acute dose for infants and children 10 mg/kg bw and for adults 25 mg/kg bw. However, alkaloids can also have medical use for their hypocholesterolemic, antiarrhythmic, and immunosuppressive activity. Bitter lupins can be detoxified by biological, chemical, or aqueous processes. The shortest debittering process requires one hour. This review presents the nutritional composition of lupins, their uses (as food, medicine, and functional protein isolates), toxicology, and debittering process scenarios. It critically evaluates the data, infers conclusions, and makes suggestions for future research.

Background: Prepared sprouts, after culturing in a medium with an increased iron concentration, could become a beneficial food iron fortificant. However, the efficient iron accumulation depends on the plants genus, species and/or varieties. The aim of the study was to indicate the seeds or grains which accumulate iron most efficiently during the sprouting process., Methods: Alfalfa, lentil, lupine and soybean seeds as well as wheat grains were sprouted in abiotic stress conditions induced by the excess of iron(II) in culture media. The tolerance of these plants to iron concentration and its accumulation in the material obtained (with FAAS method) were analyzed., Results: The smallest tolerance was noted for lentil seeds and wheat grains. Other plants developed in 25 mM solution of FeSO4. The highest accumulation of iron was observed in alfalfa sprouts. However, lupine and soybean seeds are the most recommended raw material for the production of the sprouts on an industrial scale.

The use of vegetable proteins as food ingredient is becoming increasingly important due to their high versatility and environmental acceptability. This work describes a chemical characterization and techno-functional properties (emulsifying and foaming properties) of 3 protein fractions obtained from a protein-rich novel lupin variety, AluProt-CGNA(®) . This nongenetically modified variety have a great protein content in dehulled seeds (60.6 g protein/100 g, dry matter), which is higher than soybean and other lupin varieties. A simple procedure was utilized to obtain 3 different fractions by using alkali solubilization and isoelectric precipitation. Fractions 1 and 3 were mainly composed of protein and polysaccharides (NNE), whereas fraction 2 was mainly composed by protein (97%, w/w). Fraction 3 presented interesting and potential foaming properties in comparison to the other fractions evaluated in the study. Besides, its solubility, foaming and emulsifying capacity were practically not affected by pH variations. The 3 fractions also presented good emulsion stability, reaching values above a 95%. SDS-PAGE showed that fractions 1 and 2 contained mainly conglutin α, β, and δ, but in different ratios, whereas fraction 3 contained mainly conglutin γ and albumins. The results of this work will provide better understanding for the utilization of each protein fractions as potential ingredients in food industry., (© 2016 Institute of Food Technologists®)

Broad beans (Vicia faba) and lupin seeds (Lupinus albus) are legumes rich in a wide range of compounds, which may represent a useful dietary approach for modulating the human gut microbiome. In this work, after in vitro digestion, legume samples were used as carbon sources in anaerobic batch cultures to evaluate their impact on the intestinal microbiota composition and on their metabolic products. The fermentations were monitored by a decrease in pH, generation of short chain fatty acids (SCFA) and lactate and the changes in the dynamic bacterial populations by fluorescence in situ hybridization (FISH). The total SCFA at the end of fermentation was 81.52 mM for lupin seeds and 78.41 mM for broad beans accompanied by a decrease of the pH for both legumes. The microbial groups that increased significantly (P < 0.05) were Bifidobacterium spp., Lactobacillus-Enterococcus, Atopobium, Bacteroides-Pretovella, Clostridium coccoides-Eubacterium rectale, Faecalibacterium prausnitzii and Roseburia intestinalis. This impact on the intestinal microbiota suggests that lupin seeds and broad beans may be used in the development of novel functional foods, which can be included in dietary strategies for human health promotion.

The 11S storage globulin of white lupin seeds binds to a metal affinity chromatography matrix. Two unusual stretches of contiguous histidine residues, reminiscent of the multiple histidines forming metal binding motifs, at the C-terminal end of 11S globulin acidic chains were hypothesized as candidate elements responsible for the binding capacity. To prove this, the protein was incubated with a lupin seed endopeptidase previously shown to cleave at twin arginine motifs, recurrent in the sequence region of interest. Upon incubation with this enzyme, the loss of metal binding capacity paralleled that of the anti-his-tag reactive polypeptides. The recovered small proteolytic fragment was analyzed by mass spectrometry and N-terminal sequencing and found to correspond to the 24-mer region cleaved off at twin arginine residues and containing the natural his-tag-like region. Similarly, when lupin seeds were germinated for a few days, the his-tag containing 11S globulin chain was converted to a form devoid of such region, suggesting that this mechanism is a part of the natural degradatory process of the protein. The hypothesis that the ordered and controlled dismantling of storage proteins may generate peptide fragments with potential functional roles in plant ontogenesis is presented and discussed.

The aim of this study was to determine the flavonoid profile of Lupinus mexicanus germinated seed extract (PE) and to evaluate its effect as a phytoestrogen on the morphometric parameters of CA3 hippocampal neurons of ovariectomized rats (OVX). L. mexicanus seeds, germinated for 48 h, were homogenized and macerated using an 80% ethanol solution. The extract was analyzed by HPLC/MS-MS. Thirty young Wistar strain female rats (200±10 g) were randomly distributed into four groups: sham operated (S) treated with dimethyl sulfoxide (vehicle); ovariectomized and treated with 1250 μg of PE extract (OVX-PE); ovariectomized and treated with 5 μg estradiol benzoate (OVX-EB); and ovariectomized and vehicle treated (OVX). All substances were injected subcutaneously daily for 28 days. On day 29, the animals were sacrificed, perfused, and fixed to obtain the brains for histological processing. Each brain was cut and stained with hematoxylin and eosin. The thickness of the stratum oriens (SO), the nuclear diameter, and the neuronal density were measured in the hippocampus CA3 area. Nine different flavonoids and one non-identified compound were detected. The histological analysis demonstrated that the thickness of the SO was higher in the OVX-EB and S groups than in the OVX-PE and OVX groups (p⟨0.05); in addition, the nuclear diameters of the neurons in the OVX-EB and S groups were higher compared with the other groups (p⟨0.05). The OVX group had the highest cellular density among groups (p⟨0.05). Based on our results, the PE obtained did not have beneficial effects on CA3 hippocampal neurons.

White lupin seed can be used for traditional and functional foods or as animal feed. This study aimed to support lupin breeders and production stakeholders by assessing the extent of genotypic, environmental, and genotype × environment (GE) interaction effects on seed contents of oil, tocopherols (TOC), and quinolizidine alkaloids (QA), grain yield, and seed weight of eight elite genotypes grown in two climatically contrasting Italian locations for two cropping years. On average, plants in the subcontinental climate site exhibited higher grain yield and seed size, about 8% lower oil content, and almost 85% higher QA content than those in the Mediterranean climate site. The range of genotype means was 2.97-5.14 t/ha for yield, 92-110 mg/g for oil, and 0.121-0.133 mg/g for TOC. TOC amount was largely unpredictable and featured large GE interactions that hinder its genetic improvement. Oil and alkaloid contents and seed size are more predictable and offer potential for selection.

The coffee berry borer Hypothenemus hampei is a pest that causes great economic damage to coffee grains worldwide. Because the proteins consumed are digested by aspartic proteases in the insect's midgut, the inhibition of these proteases by transferring a gene encoding an aspartic protease inhibitor from Lupinus bogotensis Benth. to coffee plants could provide a promising strategy to control this pest. Five aspartic protease inhibitors from L. bogotensis (LbAPI) were accordingly purified and characterized. The gene encoding the L. bogotensis aspartic protease inhibitor (LbAPI), with the highest inhibitory activity against H. hampei, was expressed in Escherichia coli and the purified recombinant protein (rLbAPI), with a molecular mass of 15 kDa, was subsequently assessed for its ability to inhibit the aspartic protease activity present in the H. hampei midgut in vitro, as well as its effects on the growth and development of H. hampei in vivo. The in vitro experiments showed that rLbAPI was highly effective against aspartic proteases from H. hampei guts, with a half maximal inhibitory concentration (IC50) of 2.9 μg. The in vivo experiments showed that the concentration of rLbAPI (w/w) in the artificial diet necessary to cause 50% mortality (LD50) of the larvae was 0.91%. The amino acid sequence of LbAPI had high homology (52-80%) to the seed storage proteins, vicilin and β-conglutin, suggesting that this protein was generated by evolutionary events from a β-conglutin precursor. Based on these results, LbAPI may have a dual function as storage protein, and as defense protein against H. hampei. These results provide a promising alternative to obtain a coffee plant resistant to H. hampei., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The aim of this study was to evaluate the effects of a different dietary content of sweet yellow lupine seed meal (YLM) on gastrointestinal tract development and function, the growth performance (13-18 weeks of age) and meat quality of growing-finishing turkeys. Control grower and finisher diets contained soybean meal (SBM), and in experimental diets, SBM was replaced with YLM at 6%, 12% and 18% (Groups L6, L12 and L18, respectively). The diets were formulated to be iso-nitrogenous and iso-energetic. In the first phase of feeding, YLM tended to decrease feed intake (p = 0.087) and body weight gain (BWG, p = 0.078) linearly due to significant deterioration in feed conversion ratio (FCR, p = 0.030). An opposite trend was noted in the second phase of feeding: BWG increased (p = 0.069) and FCR improved significantly (p = 0.004). Over the entire experiment, inclusion of YLM did not affect feed intake, BWG, FCR or excreta dry matter (DM) content. The highest YLM dietary level had no adverse effects on fermentation processes in the caeca. Positive changes, which were observed in turkeys fed YLM-supplemented diets, included an increased gizzard weight, a decreased pH of gizzard contents and a decreased viscosity of small intestinal digesta. The inclusion of YLM at 18% had no effect on carcass quality, the pH and colour intensity of breast meat. The highest inclusion rate of YLM (18%) in turkey diets significantly (p < 0.05) intensified the fatty flavour of meat and increased its hardness, springiness and chewiness, whereas it had no effect on the appearance, aroma and overall acceptability of breast meat. It can be concluded that YLM at the highest dietary rate used in the present experiment (18% of the diet) had no adverse effects on the growth performance or the analysed physiological parameters of turkeys.

In addition to their favourable nutritional profile, legumes also contain a range of bioactive compounds such as phenolic compounds and phytosterols which may protect against chronic diseases including cancer and cardiovascular disease. Germination of some legume seeds has been previously reported to increase the concentration of the bioactive compounds. In this study, the effect of germination of Australian Sweet Lupin (ASL) seeds for 9 days on the concentration of some bioactive compounds and the radical scavenging activity in the resulting flour was determined. The concentration of total phenolic compounds in methanolic extracts of germinated ASL flour was determined using Folin Ciocalteu reagent and phytosterols in oil extracts were analyzed by gas-liquid chromatography. The methanolic and oil extracts were also used to determine radical scavenging activity toward 2,2-diphenyl-1-picrylhydrazyl. In the methanolic extracts of germinated ASL flour, phenolic contents and the antioxidant activity were significantly increased following germination (700 and 1400 %, respectively). Analysis of the oil extracts of germinated ASL flour revealed that the concentration of phytosterols and the antioxidant activity were also increased significantly compared to ungerminated ASL flour (300 and 800 %, respectively). The relative proportion of phytosterols in germinated ASL flour was: β-sitosterol (60 %), stigmasterol (30 %) and campesterol (10 %). Germination increases the concentration of bioactive compounds and the radical scavenging activity in the germinated ASL flour.

Background: Australian sweet lupin (ASL) protein is conventionally isolated by alkaline extraction/acid precipitation, leaving a waste stream containing acid-soluble proteins (ASPs) and contaminating raffinose family oligosaccharides (RFOs). The foaming functionality of ASP isolated from ASL is not known, but ASP from another lupin species has demonstrated high foaming functionality., Results: Pre-soaking ASL kernels increased their protein/RFO ratio; however, some protein was lost by soaking. The foaming capacity of ASL protein isolated by different methods was ranked in the following order: alkaline extraction/isoelectric precipitation < direct acid extraction (novel ASP) < supernatant from isoelectric precipitation (conventional ASP) < ultrafiltered novel ASP = fresh egg white. Electrophoresis indicated enrichment of γ-conglutin and albumin peptides in ASPs and of a single peptide in the fibre residue from alkaline extraction., Conclusion: The high foaming capacity of ultrafiltered novel ASP, similar to that of fresh egg white, indicates the potential of this lupin protein as a food ingredient for foaming applications., (© 2013 Society of Chemical Industry.)

The need of quality protein in the aquaculture sector has forced the incorporation of alternative plant proteins into feeding diets. However, most plant proteins show lower digestibility levels than fish meal proteins, especially in carnivorous fishes. Manipulation of protein content by plant breeding can improve the digestibility rate of plant proteins in fish, but the identification of low digestibility proteins is essential. A reduction of low digestibility proteins will not only increase feed efficiency, but also reduce water pollution. Little is known about specific digestible protein profiles and/or molecular identification of more bioavailable plant proteins in fish diets. In this study, we identified low digestibility L. luteus seed proteins using Atlantic salmon (Salmo salar) crude digestive enzymes in an in vitro assay. Low digestibility proteins were identified by comparing SDS-PAGE banding profiles of digested and non-digested lupin seed proteins. Gel image analysis detected a major 12 kDa protein band in both lupin meal and protein isolate digested products. The 12 kDa was confirmed by 2D-PAGE gels and the extracted protein was analyzed with an ion trap mass spectrometer in tandem mass mode. The MS/MS data showed that the 12 kDa low digestibility protein was a large chain δconglutin, a common seed storage protein of yellow lupin. Comparison of the protein band profiles between lupin meal and protein isolates showed that the isolatation process did not affect the low digestibility of the 12 kDa protein.

The germination process can modify the chemical composition of nutrients in seeds, which can influence the digestibility and utilization of sprouts in animal diets compared to raw seeds. The aims of research were to provide controlled germination process of lupin seeds, monitor the changes in seed composition and determine the influence of the germination on the coefficients of standardized ileal digestibility (SID) of crude protein and amino acids in growing pigs, compared to raw lupin seeds. The seeds of two lupin species were used: yellow (RYL) (Lupinus luteus, cv. Lord) and blue (RBL) (Lupinus angustifolius, cv. Graf). Germination was provided in the dark at 24°C for 4 days. Nutritional and antinutritional compositions of raw and germinated seeds (GYL and GBL, respectively) were analysed. Digestibility study was performed on pigs with an average body weight of 25 kg, and the pigs were surgically fitted with a T-cannula in the distal ileum, with chromic oxide as an indicator. Seed germination increased the crude protein and fibre concentrations, but reduced the levels of the ether extract, nitrogen-free extracts and all amino acids in protein. The content of alkaloids and raffinose family oligosaccharides decreased in both lupin species. Germination had no positive impact (p>0.05) on the SID of crude protein and amino acids. Germination of lupin seeds negatively influenced the SID of lysine and methionine (p<0.05). The results of the research revealed a decrease in the concentrations of antinutritional factors in the sprouts of yellow and blue lupins compared to raw seeds; however, no positive effect was observed on the coefficients of the standardized ileal apparent digestibility of protein and amino acids., (© 2012 Blackwell Verlag GmbH.)

Lupin is an emerging crop worldwide due to its wide range of health benefits. In this study, a comprehensive proteome analysis was conducted using mature seed of four narrow leafed lupin cultivars, Uniharvest, Yorrel, Tanjil and Coromup, through two-dimensional gel electrophoresis followed by mass spectrometric protein sequencing. Two-dimensional gels recognised about 400 protein spots among the cultivars in the 10-100kDa molecular weight and 5.0-8.5 PI ranges. The results revealed a considerable variation of protein expression patterns with a total of 24 proteins showed differential expression among the cultivars, among which 19 were identified as β-conglutin, and 8 were identified as allergenic proteins. Most of the α, δ and γ-conglutins were showing similar expression among the cultivars. Overall, the differentially expressed proteins especially the cultivar specific proteins would be valuable markers for cultivar identification and for screening parental lines of low allergenicity in breeding process., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The occurrence of twin-arginine motifs (-R-R-) in the amino acid sequences of animal pro-proteins frequently defines the cleavage site(s) for their structural/functional maturation. No information is available on the presence and possible biological meaning of these motifs in the seed storage proteins. In this work, a novel endopeptidase activity with cleavage specificity to twin-arginine pairs has been detected in mature dry Lupinus albus seeds. The endopeptidase was tested with a number of endogenous and exogenous protein substrates, which were selected according to the presence of one or more twin-arginine residue motifs in their amino acid sequences. The observed hydrolysis patterns were limited and highly specific. Partial proteolysis led to stable polypeptide fragments that were characterized by 1- and 2-D electrophoresis. Selected polypeptides were submitted to N-terminal amino acid sequencing and mass spectrometry analyses. These approaches, supported by bioinformatic analysis of the available sequences, allowed the conclusion that the polypeptide cleavage events had occurred at the peptide bonds comprised between twin-arginine residue pairs with all tested protein substrates. The endopeptidase activity was inhibited by 4-(2-AminoEthyl)Benzene-Sulphonyl Fluoride hydrochloride (AEBSF), leupeptin, and serine proteinase protein inhibitors, while it was not affected by pepstatin, trans-Epoxysuccinyl-L-leucylamido(4-guanidino)butane (E64), and ethylenediaminetetraacetic acid (EDTA), thus qualifying the Arg-Arg cleaving enzyme as a serine endopeptidase. The structural features of storage proteins from lupin and other legume seeds strongly support the hypothesis that the occurrence of an endopeptidase activity cleaving -R-R- bonds may be functional to facilitate their degradation at germination and possibly generate polypeptide fragments with specific biological activity.

Lupin seed globulin proteins form complexes with flavonoids, predominantly apigenin C-glycosides. Enzymes typical for the gastrointestinal tract were used to hydrolyze lupin seed globulins. Release of native flavonoids as a result of the proteolysis reaction was observed. Different analytical methods such as size exclusion chromatography, HPLC-MS, and fluorescence spectroscopy (steady-state fluorescence, fluorescence anisotropy, fluorescence lifetimes) were used for a detailed characterization of this phenomenon. Flavonoids liberated from lupin globulin proteins as a result of pancreatin-catalyzed digestion were bound by γ-conglutin resistant to this enzyme. Two possible mechanisms of this interaction may be suggested: hydrogen bonding between oligosaccharide chains of glycoproteins and the sugar moieties of the flavonoid glycosides or electrostatic attraction between positively charged γ-conglutin and flavonoids partially ionized at pH 7.5.

Cardiovascular diseases (CVD) are the leading cause of death globally (World Health Organisation, 2011). Many of the risk factors for CVD are modifiable, including overweight and obesity. Numerous strategies have been proposed to fight CVD, with a special focus being placed on dietary interventions for weight management. The literature suggests that two nutrients, fiber and protein, may play significant roles in weight control and hence cardiovascular health. Increasing both protein and fiber in the diet can be difficult because popular low-carbohydrate and high-protein diets tend to have considerably low-fiber intakes (Slavin, 2005). One approach to obtain both is to develop functional foods using unique ingredients. Lupin flour is a novel food ingredient derived from the endosperm of lupin. It contains 40-45% protein, 25-30% fiber, and negligible sugar and starch (Petterson and Crosbie, 1990). Research conducted to date reveals that lupin-enriched foods, which are naturally high in protein and fiber, may have a significant effect on CVD risk factors. This review explores whether there is a role for fiber-, protein-, and lupin-enriched foods in improving cardiovascular health., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The aim of the present study was to evaluate the effect of a chronic oral γ-conglutin treatment in male Sprague-Dawley rats in which hyperglycaemia had been induced by supplying 10 % d-glucose in drinking-water. A γ-conglutin dosage of 28 mg/kg body weight was daily administered to animals for 21 d. Plasma glucose, insulin and glucose overloading were monitored. Chronic administration of glucose resulted in a statistically significant (P < 0·05) increase in fasting blood glucose (2·5-fold) and insulin (2·7-fold) v. the values recorded in control rats. Simultaneous treatment with γ-conglutin attenuated the rise in plasma glucose (1·9-fold) and insulin (1·8-fold) levels in the glucose-fed rats (P < 0·05). Fasting insulin and homeostasis model of insulin resistance were decreased by 34 and 48 % (P < 0·05), respectively, in the γ-conglutin-treated rats v. the values found in pair-fed animals. To confirm these results with a different approach, HepG2 cells, grown for 24 and 48 h in Dulbecco's minimum essential medium containing different glucose concentrations (5·5, 11·1 and 16·5 mmol/l), were exposed to 10 μmol/l γ-conglutin with or without 10 mmol/l metformin or 100 nmol/l insulin. γ-Conglutin increased glucose consumption (from 1·5- to 2·5-fold) in HepG2 cells, under all experimental conditions; this effect was more evident after 48 h incubation. Moreover, in this in vitro model, the addition of γ-conglutin potentiated the activity of insulin and metformin in cell glucose consumption. These findings extend the previous ones and suggest the potential use of lupin γ-conglutin in the control of glycaemia.

Condensed tannins (CT) from grape seeds (Vitis vinifera L.) were added to complex the protein fraction of Lupinus angustifolius seeds. Three CT/protein ratios were used: 96 mg/g (T(1)), 180 mg/g (T(2)) and 0 mg/g (T(0)). The CP losses in the rumen were assessed by the nylon-bag technique and CP intestinal digestibility (CPID) was estimated using an in vitro assay applying a three-step procedure: samples were subject to rumen degradation (in situ, 16 h) and the remaining residues were subject to the digestive enzymes of the abomasum and pancreas in vitro. A positive effect (p < 0.05) of the level of CT on the immediately soluble faction a and the insoluble degradable fraction b was observed between T(0) and T(2) . In the presence of CT the rumen degradation rate was reduced (p < 0.05) from 0.0763/h (T(0)) to 0.0443/h (T(2)). The application of CT showed a reduction (around 10% for T(1)) of effective rumen CP degradability. The CPID did not seem to be affected (p > 0.05) by the presence of CT. These findings suggest that the use of grape seed CT might have the potential to improve the efficiency of utilisation of the protein fraction from lupin seeds., (© 2010 Blackwell Verlag GmbH.)

Background and Aims: Lupin seed is referred to as an antidiabetic product in traditional medicine. Conglutin-γ, a lupin seed glycoprotein, was found to cause a significant plasma glucose reduction when orally administered to rats in glucose overload trials. Conglutin-γ was identified as being responsible for the claimed biological activity, and the aim of this work was to envisage its hypothetical insulin-mimetic cellular mechanism of action. Insulin is responsible for proteosynthesis control through IRS/AKT/P70S6k/PHAS1 pathways modulation, glucose homeostasis through PKC/Flotillin-2/caveolin-3/Cbl activation and muscle differentiation/hypertrophy via muscle-specific MHC gene transcription control., Methods and Results: To assess whether conglutin-γ modulates the same insulin-activated kinases, myoblastic C2C12 cells were incubated after 72 h of differentiation with 100 nM insulin or 0.5 mg/mL (∼10 μM) conglutin-γ. Metformin-stimulated cells were used as a positive control. The effect on the above mentioned pathways was evaluated after 5, 10, 20 and 30 min. In the control cells medium insulin, conglutin-γ and metformin were not added. We demonstrated that insulin or conglutin-γ cell stimulation resulted in the persistent activation of protein synthetic pathway kinases and increased glucose transport, glut4 translocation and muscle-specific gene transcription regulation., Conclusions: Our results indicate that conglutin-γ may regulate muscle energy metabolism, protein synthesis and MHC gene transcription through the modulation of the same insulin signalling pathway, suggesting the potential therapeutic use of this natural legume protein in the treatment of diabetes and other insulin-resistant conditions, as well as the potential conglutin-γ influence on muscle cells differentiation and regulation of muscle growth., (Copyright © 2009 Elsevier B.V. All rights reserved.)

Nigella sativa, lupin and artichoke seed oils have been investigated. The oils were subjected to detailed studies using gas chromatographic analysis (GLC) for fatty acids (FA, as methyl esters) and whole sterols (as silyl derivatives). Whereas, high pressure liquid chromatography (HPLC) was employed for determination of molecular species of triacylglycerols (TAG), four sterol lipids (free and acylated sterols, FS and AS, and free and acylated sterylglycosides, FSG and ASG, as their anthroylnitrile derivatives) as well as tocopherol patterns (T). The results showed that the three seed oils are rich in oleic and linoleic acids whereas, lupin had high linolenic acid content. It was found that the TAGs of the three oils showed some similarity with sunflower oil. Lupin oil had higher sterol content and it was very rich in campe- and β-sitosterol. Nigella sativa oil had a high content of isofucosterol, whereas artichoke oil was unique in having a high content of 5-stigma-, 7-stigma-, and avena- sterol. Concerning the FS and AS, Nigella sativa oil had the highest content, whereas artichoke oil had the highest content of FSG and ASG. Nigella sativa and lupin oils contained over 90 % γ-T while, artichoke oil comprised about 100 % α-T. It is recommended to use the three oils as healthy oils and folk medicine.

Lupin seed gamma-conglutin, orally administered to animal models, has been shown to display glucose-controlling properties. Therefore, we have addressed the study of gamma-conglutin susceptibility to proteolytic enzymes in vitro as the basis to unveil its metabolic fate in the body. Pepsin treatment at pH 2.0 and 3.0 caused extensive proteolytic breakdown, while at pH 4.0, where pepsin is minimally active, gamma-conglutin was unaffected. Aliquots of the pepsin-treated protein were further incubated with pancreatin at neutral pH. If the protein backbone was already cleaved by pepsin action, then the breakdown by pancreatin was almost complete; alternatively, pancreatin did not affect at all gamma-conglutin polypeptide chain. This was not due to an inhibitory activity of gamma-conglutin, because co-incubation with casein showed complete breakdown of the milk protein. Furthermore, gamma-conglutin was incubated with bromelain, a proteinase effective between pH 4.0 and 7.0. A sharp transition from the uncleavable to the fully cleavable form of gamma-conglutin was observed below pH 4.25. Therefore, it was concluded that (i) gamma-conglutin is resistant to proteolysis at pH greater than 4.0, likely because of a compact native conformation, (ii) an acidic pH renders the protein susceptible to proteases, suggesting the occurrence of a trans conformation, which has also been observed by circular dichroism spectral analysis, and (iii) the protein undergoes an "all or none" degradation pathway, regardless of the enzyme used.

Lupin products may be valuable as human foods because of their high protein content and potential anticholesterolemic properties. However, a small percentage of the population is allergic to lupin. In this study, we use in vitro IgE binding and mass spectrometry to identify conglutin beta, a major storage protein, as an allergen in seeds of Lupinus angustifolius and Lupinus albus. Purification of conglutin beta from L. angustifolius flour confirmed that serum IgE binds to this protein. Where IgE in sera recognized lupin proteins on Western blots, it recognized conglutin beta, suggesting this protein is a major allergen for lupin. The L. angustifolius conglutin beta allergen has been designated Lup an 1 by the International Union of Immunological Societies (IUIS) allergen nomenclature subcommittee.

The intake of lupin-based foods could imply the exposure of consumers to quinolizidine alkaloids. The objectives of this study were to assess the genetic variation among and within 11 geographic regions of Lupinus albus ecotypes, verify the quinolizidine alkaloids amount of alkaloid-poor L. albus and Lupinus angustifolius varieties, and assess the effect of two climatically contrasting Italian environments on the alkaloid content. The quantitation was performed by GC-MS, and in all samples lupanine was the most abundant quinolizidine alkaloid, followed by albine and 13alpha-hydroxylupanine for L. albus and by 13alpha-hydroxylupanine and angustifoline for L. angustifolius. Some regions tended to have a high (Azores) or low (Egypt, Near East, Maghreb) total alkaloids content, but the variation among ecotypes within regions was larger than that among regions following the estimation of variance components. Alkaloid-poor varieties tended to have higher total alkaloid contents when grown in the subcontinental climate site, exceeding in some cases the limit of 0.200 mg/g.