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Phytopathogenic fungi are known to produce several types of enzymes usually involved in plant cell wall degradation and pathogenesis. The increasing of global temperature may induce fungi, such as Lasiodiplodia theobromae ( L. theobromae ), to alter its behavior. Nonetheless, there is only limited information regarding the effect of temperature on L. theobromae production of enzymes. The need for new, thermostable enzymes, that are biotechnologically relevant, led us to investigate the effect of temperature on the production of several extracellular enzymatic activities by different L. theobromae strains. Fungi were grown at 25 °C, 30 °C and 37 °C and the enzymatic activities were detected by plate assays, quantified by spectrophotometric methods and characterized by zymography. The thermostability (25-80 °C) of the enzymes produced was also tested. Strains CAA019, CBS339.90, LA-SOL3, LA-SV1 and LA-MA-1 produced amylases, gelatinases, caseinases, cellulases, lipases, laccases, xylanases, pectinases and pectin liases. Temperature modulated the expression of the enzymes, and this effect was more visible when fungi were grown at 37 °C than at lower temperatures. Contrary to proteolytic and endoglucanolytic activities, whose highest activities were detected when fungi were grown at 30 °C, lipolytic activity was not detected at this growth temperature. Profiles of proteases and endoglucanases of fungi grown at different temperatures were characterized by zymography. Enzymes were shown to be more thermostable when fungi were grown at 30 °C. Proteases were active up to 50 °C and endoglucanases up to 70 °C. Lipases were the least stable, with activities detected up to 45 °C. The enzymatic profiles detected for L. theobromae strains tested showed to be temperature and strain-dependent, making this species a good target for biotechnological applications., Competing Interests: The authors declare that they have no competing interests.

Lipases are the industrially important biocatalysts, which are envisioned to have tremendous applications in the manufacture of a wide range of products. Their unique properties such as better stability, selectivity and substrate specificity position them as the most expansively used industrial enzymes. The research on production and applications of lipases is ever growing and there exists a need to have a latest review on the research findings of lipases. The present review aims at giving the latest and broadest overall picture of research and development on lipases by including the current studies and progressions not only in the diverse industrial application fields of lipases, but also with regard to its structure, classification and sources. Also, a special emphasis has been made on the aspects such as process optimization, modeling, and design that are very critical for further scale-up and industrial implementation. The detailed tabulations provided in each section, which are prepared by the exhaustive review of current literature covering the various aspects of lipase including its production and applications along with example case studies, will serve as the comprehensive source of current advancements in lipase research. This review will be very useful for the researchers from both industry as well as academia in promoting lipolysis as the most promising approaches to intensified, greener and sustainable processes. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:5-28, 2018., (© 2017 American Institute of Chemical Engineers.)

We have developed an enzymatic protocol to modify the composition of palmarosa essential oil by acylation of its alcohol components by three different acyl donors at various rates. The resulting modified products were characterized by qualitative and quantitative analyses by gas chromatography, and their olfactory properties were evaluated by professional perfumers. We showed that our protocol resulted in two types of modifications of the olfactory properties. The first and most obvious effect observed was the decrease of the alcohol content, with the concomitant increase of the corresponding esters, along with their fruity notes (pear, most notably). The second and less obvious effect was the expression of notes from minor components ((E)-β-ocimene, linalool, β-caryophyllene, and farnesene), originally masked by the sweet-floral-rose odor of geraniol, present in 70% in the palmarosa essential oil used, and emergence of citrus, green, spicy and clove characters in the modified products. This methodology might be considered in the future as a sustainable route to new natural ingredients for the perfumer., (Copyright © 2013 Verlag Helvetica Chimica Acta AG, Zürich.)

This study aimed to optimize the kinetic resolution of building blocks for the synthesis of β-blockers using Candida rugosa lipases, which could be potentially used to synthesize enantiomerically pure β-blockers further. Reaction mixtures were incubated in a thermostated shaker. Qualitative and quantitative analyses of the reaction mixtures were performed using chiral stationary phases and the UPLC-IT-TOF system. Of the 24 catalytic systems prepared, a system containing lipase from Candida rugosa MY, [EMIM][BF4] and toluene as a two-phase reaction medium and isopropenyl acetate as an acetylating agent was optimal. This resulted in a product with high enantiomeric purity produced via biotransformation, whose enantioselectivity was E = 67.5. Using lipases from Candida rugosa enables the enantioselective biotransformation of the β-blockers building block. The biocatalyst used, the reaction environment, and the acetylating agent significantly influence the efficiency of performer kinetic resolutions. The studies made it possible to select an optimum system, a prerequisite for obtaining a product of high enantiomeric purity. As a result of the performed biotransformation, the (S)-enantiomer of the β-blocker derivative was obtained, which can be used to further synthesize enantiomerically pure β-blockers. [ABSTRACT FROM AUTHOR]

This study investigated the antiobesity effects of black ginger extract (BGE) in high-fat diet (HFD)-induced obese mice. Mice were divided into six groups: normal diet control (NC, AIN-93G normal diet), 60% HFD control (HFD), HFD containing metformin at 250 mg/kg b.w. (Met, positive control), and HFD containing BGE at 5, 10, or 20 mg/kg b.w. for 15 weeks. BGE administration significantly prevented HFD-induced increases in weight gain, organ weight, and adipose tissue mass. Furthermore, it resulted in decreased adipogenesis and lipogenesis-related factors, including phosphorylated mitogen-activated protein kinase, peroxisome proliferator-activated receptor gamma, CCAAT/enhancer-binding proteins, sterol regulatory element-binding protein 1, phosphorylated cAMP response element-binding protein, glucose-6-phosphate dehydrogenase, fatty acid synthase, dephosphorylated ATP-citrate lyase, dephosphorylated acetyl-CoA carboxylase, and lipoprotein lipase, in white adipose tissues. Moreover, BGE administration enhanced lipolysis in white adipose tissue, as evidenced by elevated levels of adipose triglyceride lipase, phosphorylated hormone-sensitive lipase, and protein kinase A, along with reduced levels of perilipin and phosphodiesterase 3B. BGE induced thermogenesis in brown adipose tissues, as reflected by the increased expression of AMP-activated protein kinase, uncoupling protein 1, and carnitine palmitoyltransferase 1 and decreased levels of fatty acid-binding protein 4. In conclusion, this study provides comprehensive evidence supporting the antiobesity effects of BGE, elucidating the underlying molecular mechanisms involved in preventing weight gain, suppressing adipogenesis, promoting lipolysis, and stimulating thermogenesis. These findings suggest the potential therapeutic utility of BGE in combating obesity and associated metabolic disorders (KHGASP-2023-034). [ABSTRACT FROM AUTHOR]

Glycerol esterification produces a variety of valuable chemicals and has been a subject of great interest in recent years. Lipase-catalysed synthesis of acylglycerols reactions have many potential applications, including in the production of structured lipids, biofuels and pharmaceuticals. It offers several benefits over traditional chemical methods, including higher selectivity, milder reaction conditions and increased sustainability. In this systematic review, we summarize 52 primary research of lipase catalysed synthesis of acylglycerol published from 2012 to 2022. Different types of reactions employed in the synthesis of acylglycerols using various lipases are thoroughly discussed. The use of lipase in as biocatalyst in the synthesis of acylglycerols is safe for consumption and sustainable as it can reduce the utilization of toxic and hazardous organic solvents and chemicals that cause harm to human health and the environment. This review serves as a reference for the research and development of acylglycerols using biocatalysts and reactors. It could be an interest to the food and pharmaceutical industries or individuals who are interested to explore this field. Several limitations associated with the overall research on production of acylglycerols are presented in the conclusion section. [ABSTRACT FROM AUTHOR]

This study aimed at Candida rugosa lipase immobilization on a low-cost and readily available support. Among agro-industrial crops, hemp tea waste was chosen as the carrier because it provides higher immobilization performance than hemp flower and leaf wastes. Support characterization by ATR-FTIR, SEM and elemental analysis and the optimization of the adsorption immobilization process were performed. The lipase adsorption immobilization was obtained by soaking the support with hexane under mild agitation for 2 h and a successively incubating the enzyme for 1 h at room temperature without removing the solvent. The esterification of oleic acid with n-decanol was tested in a solvent-free system by studying some parameters that influence the reaction, such as the substrates molar ratio, the lipase activity/oleic acid ratio, reaction temperature and the presence/absence of molecular sieves. The biocatalyst showed the ability to bring the esterification reaction to equilibrium under 60 min and good reusability (maintaining 60 % of its original activity after three successive esterification reactions) but low conversion (21 %) at the optimized conditions (40 °C, 1:2 substrates molar ratio, 0.56 lipase/oleic acid ratio, without sieves). Comparing the results with those obtained by free lipase form at the same activity (1 U) and experimental conditions, slightly higher conversion (%) appeared for the free lipase. All this highlighted that probably the source of lipase for its carbohydrate-binding pocket and lid structure affected the esterification of oleic acid but certainly, the immobilization didn't induce any lipase conformational change also allowing the reuse of the catalytic material. • Hemp-based products to improve the sustainability of lipase immobilization. • Hemp tea waste was the best hydrophobic support for lipase adsorption. • Oleic acid was esterified by decanol in solvent-free system. • Factors affecting lipase-catalyzed esterification are described and evaluated. [ABSTRACT FROM AUTHOR]

The enzyme market is growing constantly, and lipases, in their free and immobilized forms, constitute an important part of this market. This study aimed to analyze and discuss current market scenarios using data from the literature, focusing on lipases and the feasibility of their use in the immobilized form. It also compares the cost of biodiesel production using two commercial lipases: Eversa Transform 2.0 (free lipase) and Novozyme 435 (immobilized lipase). The results show that the European and North American enzyme markets are the most prominent worldwide, with Denmark and the USA as the major exporters and importers, respectively. Lipases can be used in a wide variety of fields, and immobilization brings many advantages to this enzyme class. Finally, we show the use of immobilized enzymes and lipases in biorefinery and food industries and carry out a comparative cost analysis to produce biodiesel using a free and immobilized lipase. We concluded that the enzyme market, in which lipases play an important role, is extensive, and that immobilized lipases seem to be excellent biocatalysts with feasible costs for industrial applications. [ABSTRACT FROM AUTHOR]

With the industrialization and development of modern science, the application of enzymes as green and environmentally friendly biocatalysts in industry has been increased widely. Among them, lipase (EC. 3.1.1.3) is a very prominent biocatalyst, which has the ability to catalyze the hydrolysis and synthesis of ester compounds. Many lipases have been isolated from various sources, such as animals, plants and microorganisms, among which microbial lipase is the enzyme with the most diverse enzymatic properties and great industrial application potential. It therefore has promising applications in many industries, such as food and beverages, waste treatment, biofuels, leather, textiles, detergent formulations, ester synthesis, pharmaceuticals and medicine. Although many microbial lipases have been isolated and characterized, only some of them have been commercially exploited. In order to cope with the growing industrial demands and overcome these shortcomings to replace traditional chemical catalysts, the preparation of new lipases with thermal/acid-base stability, regioselectivity, organic solvent tolerance, high activity and yield, and reusability through excavation and modification has become a hot research topic. [ABSTRACT FROM AUTHOR]

Background: Today, biologically active additives, which contain hydrolytic enzymes (amylase, lipase, protease), are widely used in the digestive industry. These enzymes are often obtained from animal and vegetable raw materials, -for example, from the pancreas of a reindeer, the Fabricius bag of chickens, and Medusomyces Gisevii Lindau (Kombucha). However, in obtaining enzymes, the activity of amylase and lipase may decrease due to active forms --of proteases. Objective: The main goal of this research work was to evaluate the stabilizing effect of nanostructures-cyclodextrins on the activity of hydrolytic enzymes (protease, amylase, lipase) obtained from animal raw material (chicken bursa of Fabricius) and microorganism cultures (Kombucha). Identify the relationship at which the negative influence of external environmental factors is leveled and, as a result, enzymatic activity is maintained for a certain period of time. Methods: The protein concentration in the solution was determined using the Lowry method. A modified method by Rukhlyadiva was used to determine the amylolytic activity. The definition of lipolytic activity is based on modified Shibabi-Bishop method. To determine the amylolytic activity, a modified method by Rukhlyadiva was used. Results: The relationship between b-cyclodextrin and enzymes was determined, at which enzymatic activity is maintained for a certain period. Conclusion: Thus, cyclodextrins in solution stabilizes the activity of proteases, amylases, and lipases, which is of great practical importance in the food and pharmaceutical industries. [ABSTRACT FROM AUTHOR]

The current investigation was conducted with the objective of identifying yeasts that possess the ability to secrete a range of hydrolytic enzymes. Yeast isolates were collected from various samples, and the morphological characteristics of both the colony and yeast cells were examined. The capacity for enzyme production was assessed in multiple isolates by inoculating them in different media and quantifying their activities in terms of pectinase, amylase, urease, lipase, β-galactosidase, and protease. Out of a total of 42 yeast isolates, 30 displayed enzymatic activity. It is plausible that the yeast isolates obtained in this study hold potential for the production of industrial enzymes. Molecular identification techniques were employed to identify two isolates that exhibited multi-enzyme production and the highest enzyme activities. It was determined that these isolates belonged to two genera of yeasts, namely Pichia and Hanseniaspora. Specifically, isolate Y 16 was identified as Pichia kudriavizvii OL621856, while isolate Y 26 was identified as Hanseniaspora guillermondii OL621857 strains. [ABSTRACT FROM AUTHOR]

There has been a growing interest in ecofriendly enzymatic processes. However, enzyme solubility limits the application of many biocatalysts in continuous systems, requiring the development of cost‐effective strategies for enzyme immobilization. Based on this premise, this study investigated the application of lipase immobilized in starch–alginate beads for oil transesterification in a tubular reactor. An economical derivative was produced by immobilizing Eversa Transform 2.0 in 50:50 (w/w) starch–alginate beads using the jet‐cutting technique. The biocatalyst had a particle size of about 500 μm and activity of 138.67 ± 18.53 U g−1. X‐ray photoelectron spectroscopy showed nitrogen content ranging from 6.38% to 7.29%, with uniform distribution of lipase throughout the beads. Nitrogen isotherms were characteristic of mesoporous materials, with an average pore diameter of 48.09 Å and low surface area (0.69 m2 g−1). A face‐centered central composite design was used to study soybean oil transesterification. In the best four runs, the process achieved a mean triglyceride conversion of 45%. High ester productivity levels (2.05 × 10−2% ester g−1 biocatalyst min−1 or 1.5 × 10−4% ester U−1 min−1) were obtained. Biocatalyst reuse led to a twofold increase in ester concentration (14.57% vs 7.7%). These findings confirm the successful development of a low‐cost biocatalyst suitable for use in continuous reactions. [ABSTRACT FROM AUTHOR]

The lipase from Prunus dulcis almonds was inactivated under different conditions. At pH 5 and 9, enzyme stability remained similar under the different studied buffers. However, when the inactivation was performed at pH 7, there were some clear differences on enzyme stability depending on the buffer used. The enzyme was more stable in Gly than when Tris was employed for inactivation. Then, the enzyme was immobilized on methacrylate beads coated with octadecyl groups at pH 7 in the presence of Gly, Tris, phosphate and HEPES. Its activity was assayed versus triacetin and S -methyl mandelate. The biocatalyst prepared in phosphate was more active versus S -methyl mandelate, while the other ones were more active versus triacetin. The immobilized enzyme stability at pH 7 depends on the buffer used for enzyme immobilization. The buffer used in the inactivation and the substrate used determined the activity. For example, glycine was the buffer that promoted the lowest or the highest stabilities depending on the substrate used to quantify the activities. • The activity and stability of the lipase from almond depends on the buffer at pH 7. • The enzyme was immobilized via interfacial activation using different buffers at pH 7. • The properties of the biocatalysts were quite different. • The enzyme stability depended on the used substrate for activity determination. • A strong relation between inactivation and immobilization buffer and substrate was detected. [ABSTRACT FROM AUTHOR]

The proportion of fats or oils in natural or artificial feed is generally 6%-30%, since this is essential to cover fish's energetic and structural requirements. Therefore, studies of the ontogeny or response of lipase activity to food treatments are widespread. A systematic review of articles published over 5 years (2016-2020) on lipase activity in fish in aquaculture was carried out; however, this was taken only as a representative example. Any 5-year period between 201-2022 would have shown similar results in terms of the actual lipase method used. As a result of this review, it was found that the methods used by the authors are very varied and have significant differences in terms of the type of substrate, substrate concentration, bile salt type and concentration, pH, temperature, incubation time, measurement of hydrolysis products, and definition of lipase units. The above does not mean that comparison of these studies is of no value, but that it is significantly limited. The most used methods (with p-nitrophenyl derivates, β-naphthyl derivates, and emulsified natural oils as substrate) can be reviewed to determine the most appropriate standard curves or the corresponding molar extinction coefficient for defining the lipase units. Standardizing current lipase analytical procedures should improve the reliability of comparative studies of aquaculture fish species. [ABSTRACT FROM AUTHOR]

Brazil has one of the greatest biodiversities on the planet, where various crops play a strategic role in the country's economy. Among the highly appreciated biomasses is babassu, whose oil extraction generates residual babassu mesocarp (BM), which still needs new strategies for valorization. This work aimed to use BM as a support for the immobilization of Thermomyces lanuginosus lipase (TLL) in an 8.83 mL packed‐bed reactor, followed by its application as a biocatalyst for the synthesis of hexyl laurate in an integrated process. Initially, the percolation of a solution containing 5 mg of TLL at 25 °C and flows ranging from 1.767 to 0.074 mL min−1 was investigated, where at the lowest flow rate tested (residence time of 2 h), it was possible to obtain an immobilized derivative with hydrolytic activity of 504.7 U g−1 and 31.7 % of recovered activity. Subsequent studies of treatment with n‐hexane, as well as the effect of temperature on the immobilization process, were able to improve the activities of the final biocatalyst BM‐TLLF, achieving a final hydrolysis activity of 7023 U g−1 and esterification activity of 430 U ⋅ g−1 against 142 U g−1 and 113.5 U g−1 respectively presented by the commercial TLIM biocatalyst. Desorption studies showed that the TL IM has 18 mg of protein per gram of support, compared to 4.92 mg presented by BM‐TLL. Both biocatalysts were applied to synthesize hexyl laurate, achieving 98 % conversion at 40 °C within 2 h. Notably, BM‐TLLF displayed exceptional recyclability, maintaining catalytic efficiency over 12 cycles. This reflects a productivity of 180 mg of product ⋅ h−1 U−1 of the enzyme, surpassing 46 mg h−1 U−1 obtained for TLIM. These results demonstrate the efficacy of continuous flow technology in creating a competitive and integrated process offering an exciting alternative for the valorization of residual lignocellulosic biomass. [ABSTRACT FROM AUTHOR]

Biodiesel, a third-generation biofuel, offers several advantages over regular diesel fuel. Waste cooking oil (WCO) emerges as an ideal feedstock due to its availability and easy accessibility. In this work, biodiesel was produced from two different types of immobilized lipases: Rhizomucor miehei lipase (RMIM) and Candida antarctica lipase B (CALB). The objective of this study was to evaluate the impact of the molar ratio of WCO to methyl acetate, toward biodiesel yield and triacetin formation, from transesterification reaction when using two different immobilized lipases namely, Rhizomucor miehei lipase (RMIM) and Candida antarctica lipase B (CALB) as catalyst, as well as to assess its resulting biodiesel in a diesel engine. The enzymatic transesterification reaction was carried out with ultrasonic assistance, and the results showed that the greatest yield of 81.20% was achieved at 45 °C using CALB as a biocatalyst, with a lipase concentration of 1.8% (w/v) and an oil to methyl acetate molar ratio of 1:12 within 3 h. Triacetin, a by-product was determined their concentration for each molar ratio and analyzed using FTIR range of 500 cm-1 to 4000 cm-1, revealing a significant absorption peak at 1238.90 cm-1. Biodiesel was then blended with commercial diesel fuel in varying quantities of 7, 10, and 20% by volume (B20). The results were compared to Industrial Diesel Fuel 7% (B7) and Commercial Diesel Fuel 10% (B10). NOx and CO2 emission drops as the percentage of diesel/biodiesel blends increases, supporting WCO as a cost-effective biodiesel feedstock with low petrol pollution. [ABSTRACT FROM AUTHOR]

Accumulation of polyethylene terephthalate (PET) polyester in ecosystems across the globe is a major pollution of concern. Microbial degradation recently generated novel insights into the biodegradation of varieties of plastics. In this study, a PET degrading bacterium Brucella intermedia IITR130 was isolated from a contaminated lake ecosystem at Pallikaranai, Chennai, India. Incubation of the bacterium along with the PET sheet (0.1 mm thickness) for 60 days resulted in 26.06% degradation, indicating a half-life of 137.8 days. Considerable changes in the surface morphology of the PET sheet were found as holes, pits, and cracks on incubation with strain IITR130, as revealed by scanning electron microscopy (SEM). After bacterial treatment of PET, the formation of new functional groups, most notably in the area of 3326 cm−1 suggestive of O–H stretch, leading to carboxylic acid and alcohol as products were suggested by fourier transform infrared (FTIR) analysis. Monomethyl terephthalate (MMT) and terephthalic acid (TPA) were identified by gas chromatography–mass spectrometry (GC–MS) analysis as PET degradation metabolites. Tributyrin clearance assay confirmed the presence of a lipase/esterase enzyme in the strain IITR130. In this study, a degradation pathway for PET by an isolated and identified bacterium Brucella intermedia IITR130 was characterized in detail. [ABSTRACT FROM AUTHOR]

An enzymatic approach for the synthesis of intermediate alcohol of the drug tegoprazan in 99% ee at 20 °C in hexane has been developed utilizing commercially available immobilized Thermomyces lanuginosus lipase (TL lipase) as a biocatalyst and vinyl acetate as a acyl donor. This method involves the enantioselective esterification of (RS)-5,7-difluorochroman-4-ol with vinyl acetate using TL Lipase. This enzymatic approach provides the (S)-chiral alcohol in 44% yield with 99% enantiomeric excess (ee) and the (R)-acetate in 43% yield and >99% ee. It is a sustainable approach for the synthesis of enantiopure tegoprazan intermediate, contributing to the advancement of green chemistry. The recovered TL lipase was reused in 13 subsequent runs without loss of activity. [ABSTRACT FROM AUTHOR]

Carboxylesterases comprise a major class of α/β‐fold hydrolases responsible for the cleavage and formation of ester bonds. Found ubiquitously in nature, these enzymes are crucial for the metabolism of both endogenous and exogenous carboxyl esters in animals, plants and microorganisms. Beyond their essential physiological roles, carboxylesterases stand out as one of the important classes of biocatalysts for biotechnology. BlEst2, an enzyme previously classified as Bacillus licheniformis esterase, remains largely uncharacterized. In the present study, we elucidate the structural biology, molecular dynamics and biochemical features of BlEst2. Our findings reveal a canonical α/β‐hydrolase fold similar to the ESTHER block L of lipases, further augmented by two additional accessory C‐terminal domains. Notably, the catalytic domain demonstrates two insertions, which occupy conserved locations in α/β‐hydrolase proteins and commonly form the lid domain in lipase structures. Intriguingly, our in vitro cleavage of C‐terminal domains revealed the structure of the active form of BlEst2. Upon activation, BlEst2 showed a markedly elevated hydrolytic activity. This observation implies that the intramolecular C‐terminal domain serves as a regulatory intramolecular inhibitor. Interestingly, despite exhibiting esterase‐like activity, BlEst2 structural characteristics align more closely with lipases. This suggests that BlEst2 could potentially represent a previously unrecognized subgroup within the realm of carboxyl ester hydrolases. [ABSTRACT FROM AUTHOR]

The endocannabinoid system, known for its regulatory role in various physiological processes, relies on the activities of several hydrolytic enzymes, such as fatty acid amide hydrolase (FAAH), N-acylethanolamine-hydrolyzing acid amidase (NAAA), monoacylglycerol lipase (MAGL), and α/β-hydrolase domains 6 (ABHD6) and 12 (ABHD12), to maintain homeostasis. Accurate measurement of these enzymes' activities is crucial for understanding their function and for the development of potential therapeutic agents. Fluorometric assays, which offer high sensitivity, specificity, and real-time monitoring capabilities, have become essential tools in enzymatic studies. This review provides a comprehensive overview of the principles behind these assays, the various substrates and fluorophores used, and advances in assay techniques used not only for the determination of the kinetic mechanisms of enzyme reactions but also for setting up kinetic assays for the high-throughput screening of each critical enzyme involved in endocannabinoid degradation. Through this comprehensive review, we aim to highlight the strengths and limitations of current fluorometric assays and suggest future directions for improving the measurement of enzyme activity in the endocannabinoid system. [ABSTRACT FROM AUTHOR]

Immobilized lipases are biotechnological spotlights as tools to improve and enhance enzyme applicability. Here, lipase from Rhizomucor miehei (RML) was immobilized onto core/shell supports, named poly(methyl methacrylate) (PMMA/PMMA); poly(methyl methacrylate) copolymerized with divinylbenzene (PMMA- co -DVB/PMMA- co -DVB); and polystyrene copolymerized with divinylbenzene (PS- co -DVB/PS- co -DVB), and on a commercial non-core/shell porous support (polypropylene - Accurel MP 1000). Different enzyme loadings were evaluated (400 U.g−1, 500 U.g−1, 600 U.g−1, 4000 U.g−1, and 6000 U.g−1), and the immobilized biocatalysts were used in hydrolysis (using p -nitrophenyl laurate (p -NPL) and p -nitrophenyl butyrate (p -NPB) as substrates) and in esterification reactions (using oleic acid and ethanol as substrates). The influence of pH, temperature, and buffer concentration in the hydrolysis of p -NPL was also investigated. The immobilized biocatalysts were compared to free RML and/or to commercially immobilized RML (RM-IM) and/or to Novozym 435. RML interacted differently with each support, being RML-PMMA- co -DVB/PMMA- co -DVB and RML-PMMA/PMMA the biocatalysts with the highest enzymatic activities (hydrolytic activity: >400 U.g−1, using p -NPL; esterification activity: 25,532 U.g−1). Immobilization improved the thermal and pH tolerance and buffer concentration tolerance of RML. Both free RML and the immobilized biocatalysts exhibited their maximum activity at pH 7, with the exception of RML-Accurel MP 1000, which displayed optimal performance at pH 9.0. In addition, RML-PS- co -DVB/PS- co -DVB demonstrated increased activity at 70 °C. Notably, RML-PMMA/PMMA and RML-PMMA- co -DVB/PMMA- co -DVB showcased a maximum activity of 120 U/g, and this was achieved at different temperatures, 30°C and 50°C, respectively. This diverse library of immobilized biocatalysts shows promising potential for application in various industrial processes. [Display omitted] • RML was immobilized onto four home-made hydrophobic supports. • The properties of RML biocatalysts were modulated by the immobilization conditions. • The esterification performance surpassed RM-IM and Novozym 435. • The optimal conditions to RML biocatalysts depend of the support used. [ABSTRACT FROM AUTHOR]

Enzymatic degradation of polyethylene terephthalic acid (PET) has been gaining increasing importance. This has resulted in a significant increase in the search for newer enzymes and the development of more efficient enzyme-based systems. Due to the lack of a standard screening process, screening new enzymes has relied on other assays to determine the presence of esterase activity. This, in turn, has led to various nomenclatures and methods used to describe them and measure their activity. Since all PET-hydrolyzing enzymes are α/β hydrolases, they catalyze a serine nucleophilic attack and cleave an ester bond. They are lipases, esterases, cutinases and hydrolases. This has been used interchangeably, leading to difficulties while comparing results and evaluating progress. This review discusses the varied enzyme nomenclature being adapted, the different assays and analysis methods reported, and the strategies used to increase PET-hydrolyzing enzyme efficiency. A section on the various ways to quantify PET hydrolysis is also covered. [ABSTRACT FROM AUTHOR]

Biocatalytic engineering was carried out by varying monotonically the binary CNTs-silica composition and, accordingly, the physicochemical characteristics of adsorbents developed for immobilization of recombinant T. lanuginosus lipase (r Pichia /lip). The adsorbents based on composite carbon-silica materials (CCSMs) were produced by impregnating finely dispersed multi-walled carbon nanotubes with silica hydrosol followed by calcination in argon at 350°C; the mass ratio of the hydrophobic and the hydrophilic components varied over a wide range. Biocatalysts (BCs) for green low-temperature synthesis of various esters in a non-aqueous medium of organic solvents were prepared by adsorption of r Pichia /lip with subsequent drying under ambient conditions. The characteristics of the CCSMs and BCs were characterized by thermogravimetry, nitrogen porosimetry and electron microscopy. The catalytic properties of BCs, such as enzymatic activity, substrate conversion and specificity, as well we their operational stability depending on the chemical composition of CCSMs were extensively studied in the esterification of saturated monocarboxylic acids (C 4 , C 7 , C 18) and primary aliphatic alcohols (C 2 , C 4 , C 16) in hexane at 20°C. It was found that the esterifying activity manyfold decreased with increasing the silica content primarily due to a decrease in adsorption ability of CCSMs toward r Pichia /lip. The substrate specificity and operational stability of the lipase-active BCs did not greatly depend on the composition of CCSMs. Biocatalysts retained more than half of their initial esterifying activity after 10 reaction cycles. • Recombinant lipase r Pichia /lip was adsorbed on carbon-silica composite materials. • The binary composition of MWCNTs-silica materials affected the properties of the biocatalysts. • Modulation of esterifying activity, substrate specificity and operational stability were studied. • At high silica content, a multifold decrease in activity was due to decrease in lipase adsorption. • The optimal MWCNTs content in the developed composite adsorbents exceeded 90 wt%. [ABSTRACT FROM AUTHOR]

This document discusses the development of an efficient method for the kinetic resolution of ¿-allenols using a commercially available immobilized lipase, Lipozyme TL-IM. The study found that the method had high enantioselectivity, with an S-factor of over 1000. The researchers also found that the method was compatible with a wide range of substrates, including those with electron-withdrawing and electron-donating groups on the benzene ring. This method provides a valuable approach for synthesizing chiral oxycycles. [Extracted from the article]

Lipases are important biocatalysts and ubiquitous in plants, animals, and microorganisms. The high growth rates of microorganisms with low production costs have enabled the wide application of microbial lipases in detergent, food, and cosmetic industries. Herein, a novel lipase from Lacticaseibacillus rhamnosus IDCC 3201 (Lac-Rh) was isolated and its activity analyzed under a range of reaction conditions to evaluate its potential industrial application. The isolated Lac-Rh showed a molecular weight of 24 kDa and a maximum activity of 3438.5 ± 1.8 U/mg protein at 60 °C and pH 8. Additionally, Lac-Rh retained activity in alkaline conditions and in 10% v/v concentrations of organic solvents, including glycerol and acetone. Interestingly, after pre-incubation in the presence of multiple commercial detergents, Lac-Rh maintained over 80% of its activity and the stains from cotton were successfully removed under a simulated laundry setting. Overall, the purified lipase from L. rhamnosus IDCC 3201 has potential for use as a detergent in industrial applications. Key points: • A novel lipase (Lac-Rh) was isolated from Lacticaseibacillus rhamnosus IDCC 3201 • Purified Lac-Rh exhibited its highest activity at a temperature of 60 °C and a pH of 8, respectively • Lac-Rh remains stable in commercial laundry detergent and enhances washing performance [ABSTRACT FROM AUTHOR]

Additives are used to improve bread quality; for this reason, lipase stands out, as it simulates the effect of emulsifiers. The present work aimed to verify the effect of commercial lipase addition on the physical and technological characteristics of the bread. The first stage used 3³ factorial design namely three enzymes, five concentrations, and three fermentation times. The second stage used 2³ factorial design, and stored over 28 days, with two formulations (control and with enzyme) and three storage temperatures. The PCA identified the variables with the most significance in PC1: specific volume, volume, and variables related to texture and expansion. In the second stage (shelf life), the PCA revealed that the bread samples with enzymes in the formulation differed from the control samples. Based on the circle of correlations, enzyme samples were distinguished by lower chewiness, gumminess, and hardness. Microscopic analysis showed that the starch molecules were more uniform in the first 14 days. However, the starch molecules lost their conformational structure afterwards. Therefore, the incorporation of lipase improved the bread’s technological parameters such as volume, texture, and structure. [ABSTRACT FROM AUTHOR]

With the global increase in hyperglycemia and hyperlipidemia, there is an urgent need to explore dietary interventions targeting the inhibition of dipeptidyl peptidase-IV (DPP-IV) and lipid digestion and absorption. This study investigated how Lactobacillus rhamnosus GG (LGG) affects various aspects of black goji berry (BGB) (Lycium ruthenicum Murr.) juice, including changes in physicochemical and functional properties, as well as microbiological and sensory attributes. Throughout the fermentation process with 2.5–10% (w/v) BGB, significantly improved probiotic viability, lactic acid production, and decreased sugar content. While total flavonoids increase, anthocyanins decrease, with no discernible change in antioxidant activities. Metabolite profiling reveals elevated phenolic compounds post-fermentation. Regarding the inhibition of lipid digestion and absorption, fermented BGB exhibits improved bile acid binding, and disrupted cholesterol micellization by approximately threefold compared to non-fermented BGB, while also increasing pancreatic lipase inhibitory activity. Furthermore, a decrease in cholesterol uptake was observed in Caco-2 cells treated with fermented BGB (0.5 mg/mL), with a maximum reduction of 16.94%. Fermented BGB also shows more potent DPP-IV inhibition. Sensory attributes are significantly improved in fermented BGB samples. These findings highlight the potential of BGB as a bioactive resource and a promising non-dairy carrier for LGG, enhancing its anti-hyperglycemic and anti-hyperlipidemic properties. [ABSTRACT FROM AUTHOR]

Lipase is one of the most commonly used biocatalysts in the food, pharmaceutical and cosmetic industries, and can be produced by Yarrowia lipolytica yeast. Despite the intensive studies of lipase from Yarrowia, there are still many open questions regarding the enzyme secretion process, especially by new isolates of this genus as well as the effect of substrates or surfactants, or both on the production of lipase. This research focused on the effect of olive oil and surfactant Tween 80 including the optimisation of the concentration of these compounds on the production of lipase by some novel Yarrowia isolates. Moreover, the optimal environmental parameters (pH, temperature) of crude enzyme synthetised by Yarrowia strains were determined. All investigated strains were able to produce lipase in both intracellular and extracellular fractions. The extracellular lipase activities were higher than the intracellular ones (Y. divulgata Y.02062 and Yarrowia lipolytica 854/4 147 U/L, 80 U/L and 474 U/L, 122 U/L, respectively). In the case of extracellular lipase, supplementing olive oil and Tween 80 enhanced significantly the synthesis and secretion of the enzyme. The lipase activity can even be enhanced by 20 times higher from 25 U/L to 474 U/L in the case of Yarrowia lipolytica 854/4 strain. In the case of intracellular, supplementation of Tween 80 generally reduces lipase activity except for the Y. lipolytica 1/4 strain, which was affected by two times the increase. The optimised concentration of olive oil and Tween 80 were determined for Y. divulgata Y.02062, Y. divulgata 5257, Y. lipolytica 1/4, and Yarrowia lipolytica 854/4 strains as 1.6% olive oil and 0.09% Tween 80, 1.6% olive oil and 0.06% Tween 80, 1.4% olive oil and 0.09% Tween 80 as well as 1.6% olive oil and 0.065% Tween 80, respectively. The optimum pH and temperature of crude lipases (intra and extracellular) synthetised by the tested Yarrowia lipolytica and Y. divulgata yeast strains were found to be pH 7.2 and 37 °C, respectively. Our results confirmed that the new isolate Y. divulgata is a very promising species for further development for industrial use as Y. lipolytica. [ABSTRACT FROM AUTHOR]

Cryoconite is a granular structure present on the glaciers and ice sheets found in polar regions including the Himalayas. It is composed of organic and inorganic matter which absorb solar radiations and reduce ice surface albedo, therefore impacting the melting and retreat of glaciers. Though climate warming has a serious impact on Himalayan glaciers, the biodiversity of sub-glacier ecosystems is poorly understood. Moreover, cryoconite holes are unique habitats for psychrophile biodiversity hotspots in the NW Himalayas, but unfortunately, studies on the microbial diversity of such habitats remain elusive. Therefore, the current study was designed to explore the bacterial diversity of the Hamtah Glacier Himalaya using both culturable and non-culturable approaches. The culturable bacterial count ranged from 2.0 × 10³ to 8.8 × 105 colony-forming units (CFUs)/g at the different locations of the glacier. A total of 88 bacterial isolates were isolated using the culturable approach. Based on the 16S ribosomal RNA gene (16S rRNA), the identified species belong to seven genera, namely, Cryobacterium, Duganella, Janthinobacterium, Pseudomonas, Peribacillus, Psychrobacter, and Sphingomonas. In the non-culturable approach, high-throughput sequencing of 16S rRNA genes (using MiSeq) showed unique bacterial community profiles and represented 440 genera belonging to 20 phyla, namely, Proteobacteria, Actinobacteria, Firmicutes, Bacteroidetes, Chloroflexi, Acidobacteria, Planctomycetes, Cyanobacteria, Verrucomicrobia, Spirochaetes, Elusimicrobia, Armatimonadetes, Gemmatimonadetes, Deinococcus-Thermus, Nitrospirae, Chlamydiae, Chlorobi, Deferribacteres, Fusobacteria, Lentisphaerae, and others. High relative abundances of Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes were observed in the samples. Phototrophic (Cyanobacteria and Chloroflexi) and nitrifier (Nitrospirae) in bacterial populations indicated sustenance of the micro-ecosystem in the oligotrophic glacier environment. The isolates varied in their phenotypic characteristics, enzyme activities, and antibiotic sensitivity. Furthermore, the fatty acid profiles of bacterial isolates indicate the predominance of branched fatty acids. Iso-, anteiso-, unsaturated and saturated fatty acids together constituted a major proportion of the total fatty acid composition. High cold-adapted enzyme activities such as lipase and cellulase expressed by Cryobacterium arcticum (KY783365) and protease and cellulase activities by Pseudomonas sp. strains (KY783373, KY783377-79, KY783382) provide evidence of the possible applications of these organisms. Additionally, antibiotic tests indicated that most isolates were sensitive to antibiotics. In conclusion, the present study contributed for the first time to bacterial diversity and biopotentials of cryoconites of Hamtah Glacier, Himalayas. Furthermore, the cold-adapted enzymes and polyunsaturated fatty acids (PUFAs)may provide an opportunity for biotechnology in the Himalayas. Inductively coupled plasma mass spectrometry (ICPMS) analyses showed the presence of several elements in cryoconites, providing a clue for the acceleratingmelting and retreating of theHamtah glacier. [ABSTRACT FROM AUTHOR]

Yarrowia lipolytica is a robust yeast species that has gained significant attention as a biofactory for various biotechnological applications and undoubtedly can be referred to as a hidden treasure trove due to boasting a diverse array of enzymes with wide-ranging applications in multiple industries, including biofuel production, food processing, biotechnology, and pharmaceuticals. As the biotechnology field continues to expand, Y. lipolytica is poised to play a pivotal role in developing eco-friendly and economically viable bioprocesses. Its versatility and potential for large-scale production make it a promising candidate for sustainably addressing various societal and industrial needs. The current review article aimed to highlight the diverse enzymatic capabilities of Y. lipolytica and provide a detailed analysis of its relevance in biocatalysis, including the use of whole-cell catalysts and isolated enzymes. The review focused on wild-type yeast strains and their species-dependant properties and selected relevant examples of Y. lipolytica used as a host organism for overexpressing some enzymes. Furthermore, the application of Y. lipolytica's potential in enantiomers resolution, lipids processing, and biodiesel synthesis, as well as the synthesis of polymers or esterification of different substrates for upgrading biologically active compounds, was discussed. [ABSTRACT FROM AUTHOR]

Vanillin is a key organoleptic component in vanilla, one of the most important natural flavors in the industry. The limited supply of natural vanilla, extracted from cured vanilla beans, elevates the commercial value of 'natural'‐labeled vanillin, producible by microbial conversion of natural precursors such as ferulic acid, eugenol, isoeugenol, lignin, and glucose. However, cellular toxicity and undesirable overoxidation of vanillin remain challenges to the microbial methods. Here, we developed a one‐pot enzymatic conversion of capsaicinoids to vanillin by retracing the biosynthetic pathway found in Capsicum plants. The enzyme cascade consists of the hydrolysis of capsaicinoids by immobilized lipase B from Candida antarctica (CALB) and the oxidative deamination of the resulting vanillylamine by ω‐transaminase from Paracoccus denitrificans (PDTA). Process optimization of individual enzyme reactions was conducted on an analytical scale, enabling>95% reaction yield of vanillin from capsaicinoids in the one‐pot cascade reaction without undesirable accumulation of the oxidation byproduct and the inhibitory reaction intermediate, i. e. vanillic acid and vanillylamine, respectively. To verify practical applicability, we performed a preparative‐scale reaction starting with 1.75 g capsaicinoids and 2.75 g sodium pyruvate at 10 mg/mL CALB and 3 μM PDTA, leading to an 81% reaction yield and a 57% isolation yield. This study is, to the best of our knowledge, the first demonstration of one‐pot production of vanillin from capsaicinoids, which might pave the way for cost‐effective and sustainable access to natural‐labeled vanillin from a renewable feedstock readily available from Capsicum fruits. [ABSTRACT FROM AUTHOR]

Lipases are versatile biocatalysts for various biological reactions. In the detergent industry, lipases must exhibit high activity in environments with high temperature, high pH values, metal ions, and organic solvents. Therefore, researchers are intensively searching for more stable and efficient lipases. A new thermophilic lipase, LipGt6, was identified in Geobacillus thermoleovorans H9, a new thermophilic strain isolated from ultrahigh-temperature compost. A structural model of LipGt6 was constructed using an esterase from Geobacillus thermodenitrificans as a template, and site-directed mutagenesis confirmed the predicted active site residues. LipGt6 exhibited the highest activity towards medium- and long-chain fatty acids (C8–C14), and the optimum temperature and pH were 50 °C and 9.0, respectively. LipGt6 was found to be thermostable up to 70 °C. In the presence of 1% H2O2 and sodium deoxycholate, LipGt6 retained 70 to 75% relative activity. These findings reveal that LipGt6 is potentially useful for the industrial production of detergent. Based on comparison of the amino acid sequences, the enzyme belongs to a new subfamily called lipolytic enzyme family II. The catalytic residues Ser and His were more critical than Asp, and the Asp221 catalytic residue is not likely critical for the lipolytic reaction of LipGt6. [ABSTRACT FROM AUTHOR]

Lipases represent versatile biocatalysts extensively employed in transesterification reactions for ester production. Ethyl oleate holds significance in biodiesel production, serving as a sustainable alternative to petroleum-derived diesel. In this study, our goal was to prospect lipase and assess its efficacy as a biocatalyst for ethyl oleate synthesis. For quantitative analysis, a base medium supplemented with Rhodamine B, olive oil, and Tween 80 was used. Solid-state fermentation utilized crambe seeds of varying particle sizes and humidity levels as substrates. In the synthesis of ethyl oleate, molar ratios of 1:3, 1:6, and 1:9, along with a total enzymatic activity of 60 U in n-heptane, were utilized at temperatures of 30 °C, 37 °C, and 44 °C. Reactions were conducted in a shaker at 200 rpm for 60 min. As a result, we first identified Penicillium polonicum and employed the method of solid-state fermentation using crambe seeds as a substrate to produce lipase. Our findings revealed heightened lipolytic activity (22.5 Ug−1) after 96 h of fermentation using crambe cake as the substrate. Optimal results were achieved with crambe seeds at a granulometry of 0.6 mm and a fermentation medium humidity of 60%. Additionally, electron microscopy suggested the immobilization of lipase in the substrate, enabling enzyme reuse for up to 4 cycles with 100% enzymatic activity. Subsequently, we conducted applicability tests of biocatalysts for ethyl oleate synthesis, optimizing parameters such as the acid/alcohol molar ratio, temperature, and reaction time. We attained 100% conversion within 30 min at 37 °C, and our results indicated that the molar ratio proportion did not significantly influence the outcome. These findings provide a methodological alternative for the utilization of biocatalysts in ethyl oleate synthesis. [ABSTRACT FROM AUTHOR]

In this study, a Taguchi experimental design was used for optimzing the immobilization of the lipase Eversa® Transform 2.0 (EVS) onto a hybrid support consisting of chitosan (CHI) and agarose (AGA), with glutaraldehyde (GLU) used as the support activator. The biocatalyst obtained was characterized by X-Ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Thermogravimetry (TGA), Energy Dispersive Spectroscopy (EDS), and Scanning Electron Microscopy (SEM). The optimized reaction conditions (60 min, 5 mM ionic strength, 1% GLU concentration, and 5 mg protein load per g of support) resulted in a highly active biocatalyst (74.39 ± 0.48 U/g) and delivered an immobilization yield of 74.20 ± 0.28%. The biocatalyst produced was observed to lose only 15.3% of its activity after 61 days of storage. The activity was also observed to increase by 96.70% ± 0.76, 27.34% ± 2.34, and 84.35% ± 1.68 in the presence of the organic solvents hexane, cyclohexane, and methanol, respectively. Additionally, the byocatalist showed more pronounced activity at temperatures above 50 °C and was still able to retain approximately 30% of it at 70 °C. These values were found to be higher at alkaline pHs, as the activity of Eversa® 2.0 Transform saw an increase of up to 140% at pH 9. The desorption tests performed did not reveal any enzymatic detachment from the support. The novel biocatalyst also showed promising ester-lubricating properties. Furthermore, the in silico study revealed a binding affinity of −5.1 kcal/mol between oleic acid and the enzyme, suggesting that the combination of the substrate and the lipase was more stable and therefore, suitable for esterification. [ABSTRACT FROM AUTHOR]

The aim of this work, was studying the effect of cold plasma treatment on enzymes and nutritional proteins denaturation, and ocurrance of browning reactions of bottled raw milk. A surface discharge plasma system was used for this purpose. The reactor of this system was a quartz cylinder with a diameter of 1 cm and a height of 25 cm. a steel cover with a thickness of 1 mm and height of 25 cm was used on the inner surface of the reactor and as a high voltage discharge electrode. The liquid inside the bottle (milk) was also considered as neutral electrode. The time of inactivation of catalase, alkaline phosphatase, lipase, peroxidase, and protease enzymes, bovine serum albumin, immunoglobulins, alpha lactalbumin, beta lactalbumin, lysine and thiamine were investigated. The simulation was performed by COMSOL a3.5 software for a two - dimensional geometry. The results showed the deactivation time of catalase, phosphatase, and lipase is highly low while the peroxidase and protease show the longest deactivation time. However the final deactivation time of all enzymes is highly low compared with thermal treatments. The peroxidase diactivated at 0.9 min and protease deactivated at 2 minutes after plasma treatment. The other enzyme deactivation time were 0.5 seconds. Also, the protein and amino acid denaturation time has a significant difference at p< 0.05. The inactivation time of lysine amino acid was shorter than other cases studies in this work, and beta -lactalbumin protein had the longest denaturation time. Also, the time of starting the browning reaction under plasma treatment was 3.4 minutes. It can be concluded that the studied cold plasma condition have no negative effect on proteins and color of milk. [ABSTRACT FROM AUTHOR]

The slow action of fungi is one of the biggest challenges in using entomopathogenic fungi. A promising alternative to reduce the time of action is to combine conidia with extracellular enzymes. This study aimed to characterize the production of Pr1 subtilisin protease and lipases by Beauveria bassiana and Metarhizium anisopliae in different culture media and to evaluate the efficiency of the enzymatic treatment against Aphis gossypii and Spodoptera frugiperda. The isolates were cultivated in five different liquid cultures, and, after 7 days, the culture was filtered and centrifuged, and the activity of the Pr1 and lipases was measured. The fungi cultured in a Luria–Bertani broth medium had the highest activity of proteases and lipases. The mortality of A. gossypii nymphs treated with conidia 7 days after the treatment was 39% (JEF-410), 76.5% (JEF-492), 74.8% (ERL-836), and 70.9% (JEF-214). The B. bassiana JEF-410 supernatant combined with conidia increased the fungal virulence at day 5 and day 6 after treatment. When S. frugiperda larvae were treated with B. bassiana JEF-492 conidia combined with its supernatant, the time of infection was shorter compared to the larvae treated with conidia only. Once the supernatant was incubated at 37 °C, the relative activity decreased from 100% to 80% after 2 h and to 45% after 24 h. The results suggest that the supernatant of entomopathogenic fungi may be formulated and used as a biopesticide in an efficient strategy for the biological control of pests. [ABSTRACT FROM AUTHOR]

The food enzyme triacylglycerol lipase (triacylglycerol acylhydrolase, EC 3.1.1.3) is produced with the non‐genetically modified Mucor circinelloides strain AE‐LMH by Amano Enzyme Inc. A safety evaluation of this food enzyme was made previously, in which EFSA concluded that this food enzyme did not give rise to safety concerns when used in three food manufacturing processes. Subsequently, the applicant requested to extend its use to include two additional processes. In this assessment, EFSA updated the safety evaluation of this food enzyme when used in a total of five food manufacturing processes. The dietary exposure to the food enzyme‐total organic solids (TOS) was estimated to be up to 0.845 mg TOS/kg body weight (bw) per day in European populations. When combined with the no observed adverse effect level previously reported (784 mg TOS/kg bw per day, the highest dose tested), the Panel derived a margin of exposure of at least 928. Based on the data provided for the previous evaluation and the revised margin of exposure, the Panel concluded that this food enzyme does not give rise to safety concerns under the revised intended conditions of use. [ABSTRACT FROM AUTHOR]

Effects of process parameters like enzyme concentration, concentration and type of substrate, pH, temperature, speed of agitation, and pressure on lipase catalysis are reviewed. The enzyme concentration controls its interfacial presence and consequently the rate of reaction. A change in substrate concentration alters lipase kinetics. Substrate‐lipase interaction varies with substrate type and pH. Water concentration and agitation affect the extent of interfacial area. Temperature impacts the rate and thermal denaturation of enzymes. Statistical optimization can solve the problem of controlling a variable's effect by other variables. Immobilization support and nonionic surfactant altered the significance of enzyme concentration. The lipase type controlled the impact of concentrations of enzyme, substrate, and water. The water content was important during lipase‐catalyzed hydrolysis and esterification. The mode of agitation influenced the significance of enzyme concentration and temperature. Time had a remarkable impact during hydrolysis. Temperature, substrate type, and chain length notably controlled kinetic parameters. This work paves the way for similar studies on other enzymes. [ABSTRACT FROM AUTHOR]

Enzymes play an important role in numerous natural processes and are increasingly being utilized as environmentally friendly substitutes and alternatives to many common catalysts. Their essential advantages are high catalytic efficiency, substrate specificity, minimal formation of byproducts, and low energy demand. All of these benefits make enzymes highly desirable targets of academic research and industrial development. This review has the modest aim of briefly overviewing the classification, mechanism of action, basic kinetics and reaction condition effects that are common across all six enzyme classes. Special attention is devoted to immobilization strategies as the main tools to improve the resistance to environmental stress factors (temperature, pH and solvents) and prolong the catalytic lifecycle of these biocatalysts. The advantages and drawbacks of methods such as macromolecular crosslinking, solid scaffold carriers, entrapment, and surface modification (covalent and physical) are discussed and illustrated using numerous examples. Among the hundreds and possibly thousands of known and recently discovered enzymes, hydrolases and oxidoreductases are distinguished by their relative availability, stability, and wide use in synthetic applications, which include pharmaceutics, food and beverage treatments, environmental clean-up, and polymerizations. Two representatives of those groups—laccase (an oxidoreductase) and lipase (a hydrolase)—are discussed at length, including their structure, catalytic mechanism, and diverse usage. Objective representation of the current status and emerging trends are provided in the main conclusions. [ABSTRACT FROM AUTHOR]

Endophytes represent a diverse domain of microorganisms with immense biotechnological potentialities. Metabolites from endophytes, especially fungi, are useful in industrial as well as pharmaceutical aspects. Here, endophytic fungal isolates of Andrographis paniculata collected from the Tapobon, Junglemahal region of West Bengal, India, have been studied for their hydrolyzing enzyme production abilities. Out of eighty-one isolates, twenty-one were positive for amylase, protease, lipase, laccase, and tannase action. Microscopic features of positive isolates revealed that the enzyme producers were Aspergillus sp., Fusarium sp., Alternaria sp., Trichoderma sp., Exerohilum sp., Nigrospora sp., Curvularia sp., Cladosporium sp., Cochliobolus sp., Tricothecium sp., Penicillium sp., Verticillium sp., and Cephalosporium sp. The amylolytic activity was remarkable in the case of endophytic Aspergillus sp. and Fusarium sp. Also, Aspergillus sp. and Alternaria sp. had proteolytic activity. Aspergillus sp., Mycelia sterilia-2, and Trichoderma sp. were potent lipase producers. Along with Exerohilum sp. endophytic Aspergillus sp. had positive laccase activity. The tannic acid degrading activity was highest for Aspergillus sp. APL11 followed by Helicosporium sp. and Fusarium sp. Tannase producing ability of Aspergillus sp. was optimized and an incubation time-96h, incubation temperature-29°C, initial medium pH-6.5, carbon source-glucose, and nitrogen source-NaNO3 exhibited a two-fold scale-up of (2.83±0.05 u mL-1) in tannase action. These enzymes offer eco-friendly and efficient solutions across a wide range of industrial processes including food production, textile processing, and pharmaceutical synthesis. Fungal endophytes of medicinal plants act as an alternative source for hydrolysing enzymes. [ABSTRACT FROM AUTHOR]

Cold-active enzymes are extremozymes produced by psychrophiles, and have attracted much attention as biocatalysts due to their capacity to resist extreme reaction conditions in industrial processes. This study aimed to isolate bacterial strains from Arctic fjord sediments and screen them for lipase enzyme production. Out of 73 isolates, 8 were identified as good lipase producers. Bacillus cereus I13 exhibited the highest activity of 11.42 U/mL on the 4th day of incubation at 20 °C, along with the large zone of clearance of 27 mm. Optimum conditions for lipase production were identified as pH 7.0, a temperature of 20 °C, and 96 h of incubation, with glucose as a carbon source, yeast as a nitrogen source, and olive oil as a substrate. Bacillus cereus I13 has the potential to be used for the industrial production of cold-active lipase, and this study provides evidence that potent isolates can effectively increase lipase production. [ABSTRACT FROM AUTHOR]

BACKGROUND: This work aimed to study different methodologies for extracting enzymes produced by the endophytic fungi isolated from mangrove Neofusicoccum parvum, Penicillium glabra series and Bjerkandera sp. grown on organic rice. The enzymatic activities of amylase, cellulase, laccase, lipase, peroxidase, protease and keratinase were evaluated for future applications in biotechnology and enzymology. The evaluation of the extraction was carried out using the design of experiments methodology, where variations were made in pH (4–8), temperature (20–36 °C) and agitation (150–200 rpm), in times of 1, 6 and 12 h. RESULTS: Among the enzymes analyzed, promising activities were found for amylase, cellulase, lipase, protease, peroxidase and especially keratinase, presenting activities of up to 7049.47 U g−1 in the fermentation carried out with the fungus Neofusicoccum parvum, 3538.52 U g−1 with the fungus Penicillium glabra series and 5068.20 U g−1 with the fungus Bjerkandera sp. This indicates that the fungi are promising for the production of multienzyme cocktails. CONCLUSION: It should be emphasized that the microorganisms and extraction conditions evaluated resulted in a series of promising results, that is, in the case of aiming to obtain different enzymes by other extraction conditions, after fermentation of various microorganisms on rice, several options of situations are presented and validated in this work, which allows the choice of the same, depending on the future application potential of these biocatalysts. © 2023 Society of Chemical Industry (SCI). [ABSTRACT FROM AUTHOR]

The detergent formulations and the ingredients used in the formulations are variable in the entire world depending on the climate, cost, results, laundry trends and other multiple factors. The impact of detergent components on lipase (enzyme) activity was evaluated after incubation at 20 °C for 1 h and 2 h. Tween-80 demonstrated the highest residual activity, with percentages of 90.3 % and 74.6 %, respectively, inhibiting lipase activity by approximately 10 %–25 % compared to the control. Testing five laundry detergents for lipase production over various time intervals (12–16 h) and temperatures (4 °C, 20 °C, and 50 °C) indicated that "brite" detergent displayed the highest residual activity, with percentages of 84.2 %, 82.5 %, and 80.8 %, respectively, inhibiting lipase activity by approximately 15 %–20 % compared to the controls. These findings suggest that laundry detergents have a slight inhibitory effect on lipase production compared to the crude enzyme, and longer incubation periods result in decreased lipase production. [ABSTRACT FROM AUTHOR]

Utilizing covalent organic framework (COF) as a hypotoxic and porous scaffold to encapsulate enzyme (enzyme@COF) has inspired numerous interests at the intersection of chemistry, materials, and biological science. In this study, we report a convenient scheme for one‐step, aqueous‐phase synthesis of highly crystalline enzyme@COF biocatalysts. This facile approach relies on an ionic liquid (2 μL of imidazolium ionic liquid)‐mediated dynamic polymerization mechanism, which can facilitate the in situ assembly of enzyme@COF under mild conditions. This green strategy is adaptive to synthesize different biocatalysts with highly crystalline COF "exoskeleton", as well evidenced by the low‐dose cryo‐EM and other characterizations. Attributing to the rigorous sieving effect of crystalline COF pore, the hosted lipase shows non‐native selectivity for aliphatic acid hydrolysis. In addition, the highly crystalline linkage affords COF "exoskeleton" with higher photocatalytic activity for in situ production of H2O2, enabling us to construct a self‐cascading photo‐enzyme coupled reactor for pollutants degradation, with a 2.63‐fold degradation rate as the poorly crystalline photo‐enzyme reactor. This work showcases the great potentials of employing green and trace amounts of ionic liquid for one‐step synthesis of crystalline enzyme@COF biocatalysts, and emphasizes the feasibility of diversifying enzyme functions by integrating the reticular chemistry of a COF. [ABSTRACT FROM AUTHOR]

Pickering emulsions (PE) are liquid-liquid systems that are stabilized by solid (nano)particles at the fluid interface. They offer higher stability, easier separation and lower toxicity compared to classical emulsions stabilized by surfactants. Common applications range from food science to drug delivery. In the last decade they have become of more interest in the field of multiphasic biocatalysis. First, this study aims to present the influence of pH, salt strength and particle charge on enzyme activity. The different behavior of two lipases (CaLA and CRL) is shown. While the activity optimum of CaLA changed from pH 6.5 to pH 5 by applying particles with negative instead of positive surface charge, the CRL activity optimum stayed at pH 5–5.5. This enables particle charge as an additional parameter to optimize biocatalytic reactions in PEs. Second, the resulting drop sizes were measured to elucidate further interactions between the enzymes and particles in PEs. Drop sizes in PEs prepared with CaLA were not influenced by pH, but increased for positively and decreased for negatively charged particles upon the addition of CaLA. Electrostatic attraction between particles and CRL increased the droplet diameter from 10 μm up to 30 μm and therefore destabilized the PE for both particle types. • Influence of pH, salt strength and particle charge on CaLA and CRL activity inside Pickering emulsions. • activity optimum of CaLA can be shifted to a different pH with varying particle charge. • activity optimum of CRL close to IEP. • CaLA may stabilize or destabilize Pickering emulsions depending on particle charge. [ABSTRACT FROM AUTHOR]

Purpose Short bowel syndrome is a malabsorptive condition that occurs due to surgical removal or a congenital absence of a significant portion of the small intestine. Patients with short bowel syndrome often rely on parenteral support for extended periods or even their entire lives. Teduglutide, a glucagon-like peptide-2 analog, has shown promising results in reducing dependency on parenteral support in these patients by promoting intestinal adaptation and enhancing nutrient absorption. However, the long-term safety of teduglutide remains a concern, particularly with respect to its potential for the development of hyperamylasemia and hyperlipasemia. Methods This study involved patients who received teduglutide from December 2012 to December 2022 at Boston Medical Center. We evaluated outcomes and adverse events, focusing on hyperamylasemia and hyperlipasemia, through chart review. Results Thirteen eligible patients were identified who had used teduglutide. Of these, the majority (84.6%) experienced a reduction in parenteral support. A high incidence (72.7%) of nonpathological pancreatic enzyme elevation was observed in patients treated with teduglutide. These elevations were often dose dependent and were not associated with any clinical signs of acute pancreatitis or abnormal imaging findings. Conclusion This study highlights the need for further investigations into the long-term safety of teduglutide and the importance of closely monitoring amylase and lipase levels in patients undergoing treatment with teduglutide. [ABSTRACT FROM AUTHOR]

The lipases from Thermomyces lanuginosus (TLL) and Candida antarctica (B) (CALB) were immobilized on octyl-agarose beads at 1 mg/g (a loading under the capacity of the support) and by overloading the support with the enzymes. These biocatalysts were compared in their stabilities in 10 mM of sodium phosphate, HEPES, and Tris-HCl at pH 7. Lowly loaded CALB was more stable than highly loaded CALB preparation, while with TLL this effect was smaller. Phosphate was very negative for the stability of the CALB biocatalyst and moderately negative using TLL at both loadings. The stability of the enzymes in HEPES and Tris-HCl presented a different response as a function of the enzyme loading (e.g., using lowly loaded CALB, the stabilities were similar in both buffers, but it was clearly smaller in HEPES using the highly loaded biocatalysts). Moreover, the specific activity of the immobilized enzymes versus p-nitrophenol butyrate, triacetin and R- or S-methyl mandelate depended on the buffer, enzyme loading, and interaction between them. In some cases, almost twice the expected activity could be obtained using highly loaded octyl-CALB, depending on the buffer. A co-interaction between the effects on enzyme activity and the specificity of support enzyme loading and buffer nature was detected. [ABSTRACT FROM AUTHOR]

Mucor circinelloides lipase Lip10 is a dual-functional enzyme with lipolytic activity and phospholipid:diacylglycerol acyltransferase (PDAT) activity in aqueous phase. However, the catalytic properties of Lip10 and its structural characteristics remain unknown. The present study systematically investigated its PDAT activity and lipolytic activity using six phospholipid substrates, 18:0 triacylglycerol and 16:0 1,2-diacylglycerol, predict the 3D structure of Lip10 using AlphaFold2, and evaluated residues' flexibility through molecular dynamics analysis. Lip10 exhibited a preference for phosphorylethanolamine (PE) with a long-chain fatty acyl group and showed the lipolytic activity to 18:0 triacylglycerol and 16:0 1.2-diacylglycerol. 20 mM Mg2+ and Fe2+ can significantly increase the PDAT activity of Lip10 by 50–80%. Molecular docking analysis suggest that the lower binding energy for 18:0 PE with Lip10 contributes to the higher structural stability of the complex of 18:0 PE and Lip10. Mutations of Phe565Asp and Glu318Met increase PDAT activity of Lip10 by 3.6-fold and 2.7-fold, while the TAG content produced by mutants Lip10Phe103Pro, Lip10Ile487Lys and Lip10Phe570Asp decrease by 33%− 43%. The present study demonstrated structural properties of the dual-functional lipase Lip10 and provide a theoretical basis to study its roles in the lipid metabolism of M. circinelloides in the future. [Display omitted] • Lip10 is a dual-functional enzyme with lipolytic activity and PDAT activity in aqueous phase. • Lip10 exhibited stronger PDAT activity when 18:0 PE was used as the substrate. • Lip10 showed the lipolytic activity to 18:0 triacylglycerol and 16:0 1.2-diacylglycerol. • Mutations of Phe565Asp and Glu318Met increase PDAT activity of Lip10 by 3.6-fold and 2.7-fold. • TAG produced by mutants Lip10Phe103Pro, Lip10Ile487Lys and Lip10Phe570Asp decrease by 33%− 43%. [ABSTRACT FROM AUTHOR]