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Macrolactonization is a challenging process where high dilution and temperatures can extend reaction times and promote reagent degradation. Biocatalysis is a versatile strategy for synthesis but not traditionally associated within the toolbox of organic chemists for macrocyclization. Macrolactonization has been investigated using modern methods employing both catalysis and stoichiometric activation strategies on 20 different substrates with differing ring sizes and types (cyclophanes, macrolactones, macrodiolides) and structural features at the reaction site (central vs. planar chirality; primary vs. secondary alcohols). The data demonstrates that of all protocols examined, the biocatalytic route was superior, providing the highest average yields across all classes of macrocycles studied. From the stoichiometric activation strategies investigated, the Yamaguchi macrolactonization was the most versatile in terms of ring size and nature. Despite the advantages of biocatalytic macrolactonization, advances in developing supported, versatile non-enantioselective lipases would actually represent a useful tool in molecular synthesis. [ABSTRACT FROM AUTHOR]

[Display omitted] Understanding the digestion of lipid-based pharmaceutical formulations and food systems is necessary for optimising drug and nutrient delivery and has been extensively studied in bulk emulsion systems using the pH-stat method [1]. However, this approach is not suitable for investigation of individual lipid droplets, in particular the interface where the lipase acts. Microfluidic approaches to study digestion at lipid-water interfaces using droplet trapping have been proposed, however the aqueous phase in that case washes over the interface presenting uncertainty over the stoichiometry of interactions [2]. The internal interface of a Janus-like droplet, containing distinct aqueous and lipid compartments, mimics the interface of a lipid droplet in aqueous solution with controlled stoichiometry [3]. Hence, it was hypothesised that the internal interface of Janus droplets can offer a precise way to study the enzymatic digestion of lipids formulations. Using microfluidic methods, Janus-like droplets were formed by coalescing emulsion droplets containing lipid formulation and pancreatic lipase. Polarised light microscopy (PLM) and in-situ small-angle X-ray scattering (SAXS) were used to investigate the droplets. PLM revealed the growth of an aligned inverse hexagonal phase (H 2), and with SAXS showed that this phase transformation and alignment resulted from enzymatic digestion. A subsequent partial transformation from H 2 to inverse bicontinuous cubic phase occurred when simulated intestinal fluid was used instead of Tris buffer. Suggesting that phospholipids and bile salts could diffuse across the internal interface to locally affect their surroundings. [ABSTRACT FROM AUTHOR]

Introduction: Acute Pancreatitis is a medical disorder characterized by a sudden and intense inflammation of the pancreas. Patients presenting with severe acute upper abdomen discomfort should be considered for acute pancreatitis. However, to confirm the diagnosis, either biochemical or radiologic documentation is necessary. Aims and Objectives: The purpose of this study is to determine the role of biochemical marker in predicting acute pancreatitis. Methods The present research is a retrospective investigation carried out at the Department of General surgery, SCB MCH, Cuttack during the period of May 2023 to November 2023. Demographic information obtained from the medical records included age, sex, serum amylase levels, and lipase amylase lipase ratio.. Results: Out of the total of 90 patients, 73.3% (n=66) were male and 26.7% (n=24) were female. Mean age and standard deviation of patients in this research group were 38.5±2.2.5 years. The mean and standard deviation of Amylase, Lipase, and Amylase Lipase Ratio measured 870.3±754.23, 235.8±124.5, and 3.87±2.45, respectively. Amylase and Lipase exhibited sensitivity and specificity of 83.33% and 84.88% respectively, when tested three times over the normal range. The positive predictive statistic is 85.23% and the negative predictive statistic is 82.95%.Point accuracy: 84.09%. Conclusion: The ACG practice recommendations accept elevated levels of lipase and/or amylase over 3 times the upper limit of normal (ULN) as diagnostic for individuals experiencing stomach discomfort consistent with Acute Pancreatitis. Compared to amylase, lipase is somewhat more specific to Acute Pancreatitis. Elevates sooner and remains higher than serum amylase in individuals with acute pancreatitis. Diagnostic accuracy of serum lipase surpassed that of serum amylase. [ABSTRACT FROM AUTHOR]

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]

The present study aimed at synthesizing fatty acid methyl esters in a combined enzymatic method by applying degumming and transesterification of soybean oil. A soluble lipase from Serratia sp. W3 and a recombinant phosphatidylcholine‐preferring phospholipase C (PC‐PLC) from Bacillus thuringiensis were used in a consecutive manner for phosphorus removal and conversion into methyl esters. By applying 1% of recombinant PC‐PLC almost 83% of phosphorus was removed (final content of 21.01 mg/kg). Moreover, a sensitive and selective high‐performance liquid chromatography method coupled to tandem mass spectrometry was applied to obtain a comprehensive lipid profile for the simultaneous evaluation of phospholipids removal and diacylglycerol (DAG) increase. A significant increase for all the monitored DAG species, up to 138.42%, was observed by using the enzymatic degumming, in comparison to the crude sample, resulting in an increased oil yield. Serratia sp. W3 lipase was identified as a suitable biocatalyst for biodiesel production, converting efficiently the acylglycerols. The results regarding the physical‐chemical characteristics show that the cetane level, density and pour point of the obtained biodiesel are close to current regulation requirements. These findings highlight the potential of a two‐step process implementation, based on the combination of lipase and phospholipase, as a suitable alternative for biodiesel production. [ABSTRACT FROM AUTHOR]

The provision of quality and sufficient shrimp fry for production is largely determined by the management of maintenance during larval stadia. This study aims to obtain the best treatment of natural feed in the form of Spirulina sp. and Artemia sp. as feed supplements to improve the growth performance and survival rate of patin fish larvae. The experiment used a Complete Randomized Design consisting of 4 feed treatments and 3 replicates: A (Artemia sp. 100%), B (Artemia sp. 100% and Spirulina sp. 100%), C (Artemia sp. 50% and Spirulina sp. 50%), and D (Spirulina sp. 100%). The larvae with a density of 50 heads/L were kept in the aquarium and fed according to the treatment. The density of Spirulina sp. administered was 104–105 cells/mL maintained until day 7 according to the treatment. The parameters measured included growth performance, which included specific growth rate (SGR), survival (SR), absolute length growth (PPM), and analysis of larval digestive enzymes (amylase, lipase, and protease). The data were analyzed using one way-analysis of variance and continued with Duncan's Multiple Range Test with a 95% confidence interval using SPSS version 22.0. The results showed that the administration of Spirulina sp. produced better SGR, SR, PPM, amylase enzyme, lipase, and protease activities compared to the control (100% administration of Artemia sp.). The best treatment was given by 100% Spirulina sp. treatment. [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]

The study explored the chemical composition, antioxidant activity, and potential antihyperlipidemic effects of Allspice (Pimenta dioica) leaf essential oil. The oil yield from hydro-distillation was 1.61 ± 0.35% (v/w), with major compounds identified by GC-MS analysis as eugenol (72.06 ± 3.93%), β-pinene (9.29 ± 3.33%), chavicol (6.95 ± 0.6%), limonene (3.05 ± 0.71%), and 3-octenol (1.55 ± 1.07%). The oil's antioxidant activity (426 ± 3.5 mg TE g−1) was attributed to polyphenols. It also reduced porcine pancreatic lipase activity, suggesting potential in managing dietary lipids. Molecular docking showed strong binding to CES1, with eugenol, limonene, β-pinene, and octenol outperforming the control (probucol). ADME-Tox predictions indicated favorable pharmacological profiles with controlled lipophilicity and toxicity. [ABSTRACT FROM AUTHOR]

Lipases are hydrolytic enzymes which have significant potential for commercial applications, particularly in the breakdown of oil contaminants. Serratia marcescens EGHK-19 isolate exhibited considerable lipase activity. This study investigates the optimization, purification, and characterization of lipase from the Serratia marcescens EGHK-19 isolate. The optimized culture conditions revealed that maximal lipase activity was achieved after 24 hours at 30°C and pH 7, with continuous shaking at 150 rpm. Utilizing a 2% inoculum percentage with 1% diesel and 0.3% tryptone in the presence of Fe2+, Ca2+, Mg2+ salts, and Tween 80 resulted in the highest activity at 17.278 U/ml/min. The purification process involved acetone precipitation and DEAE-Sephadex column chromatography, revealing a molecular weight of approximately 60 kDa on SDS-PAGE. This method exhibited a 0.985-fold purification, and the final yield was limited to 2.097% due to lipase aggregates. Characterization of the purified lipase indicated optimal activity (8.765 U/mL/min) at 40°C a nd p H 7. T he K m a nd V max values w ere c alculated a s 6 .89 m M a nd 6 5.79 µmol/min, respectively. The presence of SDS, Tween 80, and Triton X-100 surfactants resulted in the inhibition of lipase activity. Despite these insuggestss, the biochemical characteristics of the purified lipase suggest its potential as an excellent candidate for various industrial applications. [ABSTRACT FROM AUTHOR]

In order to clarify the inhibition of total flavonoids purified extract (TFSE) of sea buckthorn peel residue on pancreatic lipase, this research optimized the purification condition of total flavonoids crude extract (TFCE) of sea buckthorn peel residue using AB-8 macroporous resin by means of static and dynamic adsorption and desorption experiment. Ultra-performance liquid chromatography-high resolution mass spectrometry (UPLC-Q-MS) was used to analyze the component of TFSE. In addition, the inhibitory effect and types of TFSE on pancreatic lipase were explored through the method of nitrophenyl butyrate combined with enzymatic reaction kinetics. The results showed that the optimal purification conditions for AB-8 macroporous resin were a sample concentration of 2.2 mg/mL, a sample pH of 5.0, a sample volume of 140 mL, and a sample flow rate of 1.0 mL/min. The volume fraction of ethanol was 75% with the eluent being 120 mL and the elution flow rate being 1.5 mL/min. After separation and purification under these conditions, the purity of TFSE increased from 9.786% to 50.204%. The total of 39 phenolic compounds were identified from TFSE with flavonoids like rutin, catechins, and quercetin-3-O-glucoside being the main compounds. TFSE had a strong inhibition on pancreatic lipase with the IC50 being 12.46 mg/mL. In addition, the inhibition type was determined to be non-competitive inhibition by using double reciprocal plot method of Lineweaver-Burk with an inhibition constant Ki being 10.21 mg/mL. TFSE has better inhibitory activity on pancreatic lipase, which is of great significance for improving the added value of sea buckthorn peel residue products. [ABSTRACT FROM AUTHOR]

BACKGROUND AND OBJECTIVES: Lipid-degrading bacteria found in processing ponds of palm oil mill effluent are recognized for the capacity to break down lipid using lipase enzyme. Identifying novel strains of these bacteria with high bioremediation potential contributes valuable insights to the sustainable management of palm oil mill effluent. Therefore, this study aimed to identify potential bacteria, assess the in vitro lipid-degrading capabilities, characterize the traits, and evaluate lipid degradation activity of potential isolates from palm oil industry wastewater. METHODS: The method used for exploring the potential of lipid-degrading bacteria in palm oil mill effluent entailed a survey comprising various stages including detection of bacteria presence, in vitro assessment of potential indices, characterization, lipid degradation testing, and determination of lipase activity. FINDINGS: The results showed that several bacteria groups were present in palm oil mill effluent, including 50-74 percent lipolytic, 31-90 percent fermentative, 76-83 percent proteolytic, and 51-74 percent cellulolytic. Selected lipid-degrading isolates demonstrated significant in vitro potential, as evidenced by high lipolytic and fermentative indices. Isolate Enzymatic 3 had the highest lipolytic index, degradation value (48.72 percent), and lipase activity (0.12 units/milliliter), identified as Bacillus cereus central carbon metabolism 2010. Similarly, isolate Fermentative 2 was found to have the highest fermentative index, degradation value (22.35 percent), and lipase activity (0.01 units/milliliter), identified as Bacillus thuringiensis American type culture collection 10792. CONCLUSION: Based on the results, isolates Enzymatic 3 and Fermentative 2 showed promising potential as biological agents for bioremediation of palm oil mill effluent. The results underscored the promising potential of specific bacteria isolates in mitigating lipidrich effluents, advocating for the integration into sustainable wastewater management practices in palm oil industry. This study provided valuable insights for future investigations aimed at unraveling the intricate mechanisms governing lipid degradation and fostering environmentally friendly solutions for industrial waste management. [ABSTRACT FROM AUTHOR]

This article discusses the loss of de novo fatty acid biosynthetic capacity in various fungal species that have developed obligate relationships with their hosts. Some fungi have expanded their metabolic pathways to enhance their fitness in specific niches, while others have downsized their core metabolic network. Certain fungal pathogens, such as Pneumocystis, Malassezia, Microsporidia, and arbuscular mycorrhizal fungi (AMF), have lost the ability to synthesize fatty acids and instead scavenge them from their hosts. The article explores the mechanisms by which these fungi assimilate fatty acids and highlights the challenges in studying and culturing these species. [Extracted from the article]

This article provides an overview of various methods for calculating free energy in the interactions between biomolecules and nanomaterials. The authors discuss the advantages and limitations of these methods and their applicability to different systems. The article aims to guide future research in this field and emphasizes the need for further improvements in force fields, software, and the use of machine learning. The document also includes a list of references to scientific articles related to drug discovery, computational methods, and the calculation of free energy in molecular systems. These articles provide valuable insights and techniques for researchers in these fields. Additionally, there are references to two specific articles that discuss the use of machine learning in drug discovery and the computational characterization of membrane proteins as potential targets for anticancer treatments. [Extracted from the article]

Alpha beta hydrolase domain containing 5 (ABHD5) is an essential coactivator of adipose triglyceride lipase (ATGL), a rate-limiting enzyme in various cell types that promotes the hydrolysis of triacylglycerol (TG) into diacylglycerol (DG) and fatty acid (FA). It acts as a critical regulatory factor in cellular lipid metabolism. The reprogramming of lipid metabolism is one of the hallmarks of cancer, suggesting that altering lipid metabolism could become a new strategy for tumor treatment. Research has revealed a close association between ABHD5 and the development and progression of malignancies. This review summarizes the role of ABHD5 in various malignant tumors and explores the different signaling pathways and metabolic routes that may be involved, providing a comprehensive mechanistic understanding of ABHD5. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the potential of ethanolic extract of propolis on the secondary lesions of the liver, renal, and pancreatic that were derived by primary colorectal cancer, and comparison of the ethanolic extract of propolis with the vitamin E. The groups included the control, ethanolic extract of propolis, vitamin E, dimethylhydrazine, dimethylhydrazine + ethanolic extract of propolis, and dimethylhydrazine + vitamin E. After 13 weeks of treatment, the blood and tissue samples were taken from all the rats, and alanine transaminase, aspartate transaminase, alkaline phosphatase, uric acid, creatinine, blood urea nitrogen, insulin, amylase, and lipase indices along with the tissue pathological examination of the kidney, liver, and pancreas were evaluated. Ethanolic extract of propolis effectively alleviated the colorectal cancer‐induced secondary lesions in the liver by significantly lowering the alanine transaminase significantly. Ethanolic extract of propolis significantly decreased uric acid in rats; and also significantly elevated the pancreatic insulin. In addition, inflammation and cell necrosis indices in all these tissues were significantly reduced when ethanolic extract of propolis was consumed compared to the dimethylhydrazine group. It seemed ethanolic extract of propolis showed high antioxidant, anticancer, and anti‐inflammatory potentials, and can be used practically to reduce the side lesions of colorectal cancer. [ABSTRACT FROM AUTHOR]

Simple Summary: This study was conducted to explore the optimal level of Rheum palmatum root (RP) for the mitigation of enteric methane and demonstrate the effects on methane production and rumen fermentation characteristics in cattle fed with RP. In the in vitro trial, 3% of RP addition most effectively reduced methane production, which determined the feeding level for the test subjects in the in vivo trial. In the in vivo trial, RP addition did not affect methane emissions in the steers, likely due to the adaptation of ruminal microorganisms to the RP addition. This study investigated the impact of Rheum palmatum root (RP) for reducing methane and its impact on rumen fermentation and blood metabolites in cattle. Rumen fluid was collected from three cannulated steers (736 ± 15 kg) and mixed with buffer (1:3 ratio) for the in vitro trial. Treatments were divided into control and RP supplement groups (1%, 3%, and 5% of substrates), with each sample incubated at 39 °C for 24 and 48 hours. Methane was measured after incubation, showing a dose-dependent linear decrease after 48 hours. Quadratic changes were observed in total volatile fatty acids, acetate, and butyrate. Additionally, in vitro dry matter digestibility decreased linearly with RP inclusion. In vivo trials involved four Korean steers in a 2 × 2 crossover design over 3 weeks, with treatments including a control group and a group with 3% RP addition. Dry matter intake (DMI) tended to decrease in the RP group compared to the control. Methane emissions (g/kg DMI) were not affected by RP addition. Blood metabolites indicated higher lipase concentrations in the RP group. In conclusion, RP reduced methane production in the in vitro trial but had no effect in the in vivo trial, likely due to adaptation of ruminal bacteria to RP. [ABSTRACT FROM AUTHOR]

A highly regioselective and efficient method has been developed for synthesizing novel β-amino alcohols fused with dihydropyrimidin-2-one. This method utilizes the enzyme Novozyme-435 to catalyze the reaction between epoxides and various aliphatic amines in acetonitrile. Novozyme-435 outperformed other catalysts, including Porcine Pancreatic Lipase (PPL), Pseudomonas aeruginosa lipase (PAL), and Candida rugosa lipase (CRL). This process yielded two series of β-amino alcohols (compounds 8a-h and 9a-h), whose structures were confirmed through IR, NMR (1H,13C), and HRMS analyses. The anti-inflammatory and antioxidant properties of these compounds were evaluated, revealing mild to moderate inhibition of TNF-α-induced ICAM-1 expression in primary human endothelial cells, with compounds 9a and 9c showing approximately 60% inhibition. Antioxidant activity, assessed using the DPPH (2,2-diphenyl-1-picrylhydrazyl) method, indicated that compounds 9a, 9b, 9c, and 9 g had the superior activity than others. This study highlights the potential of these β-amino alcohols fused with dihydropyrimidin-2-one as anti-inflammatory and antioxidant agents. [ABSTRACT FROM AUTHOR]

AbstractBiodegradation of plastic is the most eco-friendly degradation process conferred by various microbial enzymes, especially the lipase enzymes. The lipase enzymes depict a broad substrate range, high regio/stereoselectivity, and stability in organic solvents. However, enhancing the catalytic activity of lipases for industrial production remains a challenge, predominantly due to the need for their extensive mechanistic characterization. Here, we employed a multidisciplinary approach to illustrate the comprehensive physiochemical and mechanical properties of 102 lipase sequences from six different groups of plastic-degrading microorganisms. Molecular docking-based evaluation of the binding efficiencies of selected lipases with the most common plastic polymers- polyurethane (PUR) and polyethylene terephthalate (PET), revealed that the archaeal lipase had the lowest binding energy of −7.1 kcal/mol with PET. Remarkably, the molecular dynamic simulation studies (up to 100 ns) and very low RMSD (0.3 nm) and RMSF (0.25 nm) values further ascertained the stability of archaeal lipase after binding with PET polymer. Further, a 2.19 nm radius of gyration (Rg) within 100 ns simulation time, binding free energy ΔGTOTAL with −24.36 kcal/mol and entropy factor -TΔS 7.65 kcal/mol also indicated strong binding of archaeal lipase with PET polymer. The mechanistic exploration of archaeal lipase yielded valuable insights into its effective binding with PET polymer, paving the way for future lipase enzyme engineering and harnessing its industrial potential for plastic biodegradation. [ABSTRACT FROM AUTHOR]

AbstractThis study explores a novel enzymatic pretreatment approach in anaerobic reactors for dairy wastewater, using lipase AY Amano to enhance methane production and modify microbial and archaeal community composition. Batch and semi-batch reactors with a total volume of 2000 mL were used to treat dairy wastewater with initial COD of 2000 and 15,000 mg L−1, respectively. In a new novel approach, the semi-batch reactors underwent a three-phase operation: 30 days of acclimation, 30 days of rest, and 30 days of active operation. Adding lipase (0.05% wv−1) as a pretreatment significantly increased methane yield over the 90 days by 135–138% compared with the control (without enzyme addition). The organic loading rate reached 0.22 g COD day−1 L−1. Furthermore, 30 days after the end of the semi-batch reactor approach (120 days from the start), reusing sludge in batch reactors increased methane yield by 114–122% compared to the control. This increase was linked to the emergence and shift of new methanogenic communities within the sludge. Integrating hydrolytic enzymes into the anaerobic treatment enhances performance and sustainability by fostering methanogen-enriched microbial communities. This is crucial for maximizing methane production but may increase costs, requiring further economic feasibility research. [ABSTRACT FROM AUTHOR]

Backgrounds Results Conclusions Glycerolysis, with its advantages of readily available raw materials, simple operation, and mild reaction conditions, is a primary method for producing diacylglycerol (DAG). However, enzymatic glycerolysis faces challenges such as high enzyme costs, low reuse efficiency, and poor stability. The study aims to develop a cost‐effective immobilized enzyme by covalently binding lipase to pre‐activated carriers through the selection of suitable lipases, carriers, and activating agents. The optimization is intended to improve the glycerolysis reaction for efficient DAG production.Lipase CN‐TL (from Thermomyces lanuginosus) was selected through glycerolysis reaction and molecular docking to catalyze the glycerolysis reaction. Optimizing the immobilization method by covalently binding CN‐TL to poly(ethylene glycol) diglycidyl ether (PEGDGE)‐preactivated resin LX‐201A resulted in the preparation of the immobilized enzyme TL‐PEGDGE‐LX. The immobilized enzyme retained over 90% of its initial activity after five consecutive reactions, demonstrating excellent reusability. The DAG content in the product remained at 84.8% of its initial level, further highlighting the enzyme's potential for reusability and its promising applications in the food and oil industries.The immobilized lipase TL‐PEGDGE‐LX, created by covalently immobilizing lipase CN‐TL on PEGDGE‐preactivated carriers, demonstrated broad applicability and excellent reusability. This approach offers an economical and convenient immobilization strategy for the enzymatic glycerolysis production of DAG. © 2024 Society of Chemical Industry. [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]

Bioaromas can be produced by lipases either through their hydrolytic or (trans)esterifying activities. Therefore, this work reports the development of a lipase-catalyzed biotransformed licuri oil, forming volatile ethyl esters with odor notes resembling tropical fruits. Ethyl octanoate formation was promoted when 7.0 % (m/v) Lipozyme 435® was used to convert a grain alcohol:licuri oil mixture (51:49, v/v) at 58ºC and 70 rpm for 6 hours. The biotransformed oil has shown antimicrobial activity against Staphylococcus hominis , S. epidermidis, and Corynebacterium xerosis , bacteria associated with bad skin odor. Finally, this biotransformed oil was used without further treatments (e.g., recovery or purification procedures) to prepare two cosmetic formulations (in a dosage of 1.5 %), aiming for both fragrant and deodorant activity. • Odor-active ethyl esters were formed in lipase-biotransformed licuri oil. • 8.22 g/L ethyl octanoate was formed with 7 % lipase, 51:49 alcohol:oil ratio at 58°C. • Antibacterial activity of biotransformed oil could prevent bad skin odor. • Cosmetic formulations were prepared with 1.5 % biotransformed licuri oil. • No purification procedures were needed to use a natural fragrant mixture. [ABSTRACT FROM AUTHOR]

Catalytic esterification in water has been achieved with the aid of a commercially available polymer-supported lipase, Novozym-435. Remarkably, this well-known biocatalyst demonstrates significant activity towards esterification using just water as the reaction medium. This methodology features representative esterifications of complex acids and alcohols, in the presence of unprotected amines. The work disclosed leading to esters typically requires only two equivalents of alcohol, although conversion for water-soluble alcohols appears to require increased loadings. Recycling of both the aqueous medium and catalyst are documented, highlighting the potential of this new technology, especially using directed evolution on the enzyme involved. Pharmaceutically relevant compounds are efficiently esterified (e.g., Ibuprofen, Tolmetin, and Ticagrelor) and multi-step, one-pot chemoenzymatic sequences can be performed to demonstrate the robustness of this catalytic aqueous system. [ABSTRACT FROM AUTHOR]

Obesity is a condition caused by abnormal fat accumulation. The prevalence of obesity and related diseases is increasing globally, and efforts to find solutions are ongoing. In addition to diet, exercise, and surgery, drug therapy is recommended for individuals seeking a solution. Current anti‐obesity drugs target central and peripheral pathways. However, drugs targeting the central nervous system have serious side effects. Therefore, one of the approaches in the treatment of obesity is to inhibit pancreatic lipase (PL), which acts on peripheral pathways. Currently, the only clinically approved PL inhibitor is Orlistat, but this semi‐synthetic drug has also been associated with unwanted side effects. Recently, there has been extensive research on natural sources and natural products derived from these sources as potential lipase inhibitors with fewer side effects. This article reviews recent in vitro studies on inhibiting the PL enzyme by extracts from plants, fungi, and algae, as well as natural products derived from these extracts. In addition, the review presents recent molecular docking and in vivo studies demonstrating potential natural PL inhibitors. [ABSTRACT FROM AUTHOR]

Effective enzyme stabilization through immobilization is essential for the functional usage of enzymatic reactions. We propose a new method for synthesizing elastic hydroxyapatite microgel (E-HAp-M) materials and immobilizing lipase using this mesoporous mineral via the ship-in-a-bottle-neck strategy. The physicochemical parameters of E-HAp-M were thoroughly studied, revealing that E-HAp-M provides efficient space for enzyme immobilization. As a model enzyme, lipase (LP) was entrapped and then cross-linked enzyme structure, preventing leaching from mesopores, resulting in highly active and stable LP/E-HAp-M composites. By comparing LP activity under different temperature and pH conditions, it was observed that the cross-linked LP exhibited improved thermal stability and pH resistance compared to the free enzyme. In addition, they demonstrated a 156% increase in catalytic activity compared with free LP in hydrolysis reactions at room temperature. The immobilized LP maintained 45% of its initial activity after 10 cycles of recycling and remained stable for over 160 days. This report presents the first demonstration of a stabilized cross-linked LP in E-HAp-M, suggesting its potential application in enzyme-catalyzed processes within biocatalysis technology. [ABSTRACT FROM AUTHOR]

Introduction: Acute pancreatitis (AP) following drug-induced hypertriglyceridemia is a rare clinical phenomenon. Immune checkpoint inhibitors have revolutionized treatment for a variety of solid organ and hematological malignancies. Pembrolizumab is a programmed cell death receptor-1 (PD-1) inhibitor that has shown promising responses in many advanced cancers. However, a constellation of immune-related adverse events has also been described. There are reports of pembrolizumab-induced hypertriglyceridemia, but AP as a result of this side effect remains an exceedingly rare clinical sequela. Case report: We delineate a case of a patient with stage IVB non-small-cell lung cancer who developed progressive abdominal pain and nausea following administration of pembrolizumab for four months. Laboratory studies revealed increased serum lipase and triglyceride levels at 12,562 IU/L and 16,901 mg/dL, respectively. The diagnosis of AP was made based on the revised Atlanta classification criteria. After ruling out alternative causes, pembrolizumab-induced hypertriglyceridemia was considered the likely etiology of AP. Management and outcome: The patient was transferred to the medical intensive care unit for close monitoring. Treatment was initiated with intravenous fluids, pain medications, and an insulin infusion. However, her hypertriglyceridemia levels remained persistently elevated, necessitating therapeutic apheresis. She recovered well with no complications after triglyceride apheresis. Discussion: AP following pembrolizumab-associated hypertriglyceridemia remains a rare clinicopathologic entity. Given the widespread clinical use of immune checkpoint inhibitors, knowledge of such rare adverse events is crucial. Evaluation of serum triglyceride levels before and after initiating pembrolizumab therapy may be mandated, especially in patients with metabolic comorbidities. [ABSTRACT FROM AUTHOR]

Tequila vinasse, a byproduct of the tequila industry, is frequently discharged into water bodies or agricultural fields, posing significant ecological and human health risks. Bacterial communities that inhabit these agricultural fields have developed mechanisms to utilize tequila vinasse as a potential nutrient source and to promote plant growth. In this study, strains from the phyla Actinomycetota and Pseudomonadota were isolated from agricultural fields irrigated with tequila vinasse for 2, 10, and 14 years in Jalisco, Mexico. The results showed that strains of Terrabacter, Azotobacter, Agromyces, Prescottella, and Leifsonia tolerate high concentrations of tequila vinasse and promote maize seedling growth in the presence of tequila vinasse. Additionally, some of the strains solubilize potassium and produce siderophores, cellulase, protease, lipase, and esterase. The strains Terrabacter sp. WCNS1C, Azotobacter sp. WCNS1D, and Azotobacter sp. WCNS2A have potential applications in the bioremediation of tequila vinasse in agricultural fields discharged with tequila vinasse. [ABSTRACT FROM AUTHOR]

Citronellyl acetate is a crucial component of flavor and fragrance in the food, cosmetic, and pharmaceutical sectors. In non-aqueous media, it can be successfully produced via lipase catalysis. This study focuses on the lipase-catalyzed trans-esterification of citronellol alcohol with geranyl acetate ester in a non-aqueous medium to produce citronellyl acetate. For the synthesis of citronellyl acetate, crude acetone powders isolated from several plant seedlings of black cumin, fenugreek, coriander, flax, and rape seed were examined for lipase activity. Black cumin seedling lipase had the highest level of citronella acetate production with a yield of 76.32% in 72 h of reaction time. To assess the impact of different reaction parameters on citronellyl acetate production in organic solvents, hexane was chosen as the best solvent, and black cumin seedling lipase was selected as the best biocatalyst. The highest conversion yield of ester (76.32%) was found when 0.25 M of geraniol acetate and 0.25 M of citronellol reacted at 41 oC after 72 h in the presence of 0.25 g of seedling lipase enzyme in n-hexane. It has been determined that crude black cumin seedling lipase is inexpensive yet effective and has the potential to be used industrially for the synthesis of terpene esters. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus infections present a significant threat to the global healthcare system. The increasing resistance to existing antibiotics and their limited efficacy underscores the urgent need to identify new antibacterial agents with low toxicity to effectively combat various S. aureus infections. Hence, in this study, we have screened T-muurolol for possible interactions with several S. aureus-specific bacterial proteins to establish its potential as an alternative antibacterial agent. Based on its binding affinity and interactions with amino acids, T-muurolol was identified as a potential inhibitor of S. aureus lipase, dihydrofolate reductase, penicillin-binding protein 2a, D-Ala:D-Ala ligase, and ribosome protection proteins tetracycline resistance determinant (RPP TetM), which indicates its potentiality against S. aureus and its multi-drug-resistant strains. Also, T-muurolol exhibited good antioxidant and anti-inflammatory activity by showing strong binding interactions with flavin adenine dinucleotide (FAD)-dependent nicotinamide adenine dinucleotide phosphate (NAD(P)H) oxidase, and cyclooxygenase-2. Consequently, molecular dynamics (MD) simulation and recalculating binding free energies elucidated its binding interaction stability with targeted proteins. Furthermore, quantum chemical structure analysis based on density functional theory (DFT) depicted a higher energy gap between the highest occupied molecular orbital and lowest unoccupied molecular orbital (EHOMO-LUMO) with a lower chemical potential index, and moderate electrophilicity suggests its chemical hardness and stability and less polarizability and reactivity. Additionally, pharmacological parameters based on ADMET, Lipinski's rules, and bioactivity score validated it as a promising drug candidate with high activity toward ion channel modulators, nuclear receptor ligands, and enzyme inhibitors. In conclusion, the current findings suggest T-muurolol as a promising alternative antibacterial agent that might be a potential phytochemical-based drug against S. aureus. This study also suggests further clinical research before human application. [ABSTRACT FROM AUTHOR]

In order to explore the response mechanism of Lilium pumilum (L. pumilum) to saline–alkali stress, we successfully cloned LpGDSL (GDSL lipase, Gly-Asp-Ser-Leu) from L. pumilum. The qRT-PCR results indicated that the LpGDSL expression was higher in the leaves of L. pumilum, and the expression of the LpGDSL reached the highest level at 12 h in leaves under 11 mM H2O2, 200 mM NaCl, 25 mM Na2CO3, and 20 mM NaHCO3. The bacteriophage overexpressing LpGDSL was more tolerant than the control under different NaHCO3 contents. Overexpressed and wild-type plants were analyzed for phenotype, chlorophyll content, O2− content, H2O2 content, lignin content, and so on. Overexpressed plants had significantly higher resistance than the wild type and were less susceptible to saline–alkali stress. The yeast two-hybrid and BiFC assays demonstrated the existence of an interaction between LpGDSL and LpBCP. The yeast one-hybrid assay and transcriptional activation assay confirmed that B3 transcription factors could act on LpGDSL promoters. Under saline–alkali stress, L. pumilum will promote the expression of LpGDSL, which will then promotes the accumulation of lignin and the scavenging of reactive oxygen species (ROS) to reduce its damage, thus improving the saline–alkali resistance of the plant. [ABSTRACT FROM AUTHOR]

In the glycerolysis process for diacylglycerol (DAG) preparation, free lipases suffer from poor stability and the inability to be reused. To address this, a cost-effective immobilized lipase preparation was developed by cross-linking macroporous resin with poly (ethylene glycol) diglycidyl ether (PEGDGE) followed by lipase adsorption. The selected immobilization conditions were identified as pH 7.0, 35 °C, cross-linking agent concentration 2.0%, cross-linking time 4 h, lipase amount 5 mg/g of support, and adsorption time 4 h. Enzymatic properties of the immobilized lipase were analyzed, revealing enhanced pH stability, thermal stability, storage stability, and operational stability post-immobilization. The conditions for immobilized enzyme-catalyzed glycerolysis to produce DAG were selected, demonstrating the broad applicability of the immobilized lipase. The immobilized lipase catalyzed glycerolysis reactions using various oils as substrates, with DAG content in the products ranging between 35 and 45%, demonstrating broad applicability. Additionally, the changes during the repeated use of the immobilized lipase were characterized, showing that mechanical damage, lipase leakage, and alterations in the secondary structure of the lipase protein contributed to the decline in catalytic activity over time. These findings provide valuable insights for the industrial application of lipase. [ABSTRACT FROM AUTHOR]

Polyhydroxyalkanoates (PHAs) constitute a principal group of bio-degradable polymers that are produced by certain microbes under limited supply of nutrients. PHA is a linear polyester that comprises of 3-hydroxy fatty acid monomers. Triacylglycerol acylhydrolases are known to catalyze the hydrolysis of ester linkages and in turn they are beneficial in the degradation of PHA. In present study, lipase-catalyzed degradation of PHA synthesized by Priestia megatarium POD1 was monitored. A gene from thermotolerant Bacillus subtilis TTP-06 that was capable of expressing lipase enzyme was amplified by PCR, cloned into a pTZ57R/T-vector, transferred to an expression vector pET-23a (+) and expressed in Escherichia coli BL21 (DE3) cells. The recombinant enzyme purified to 19.37-fold had a molecular weight of 30 kDa (SDS-PAGE analysis). Scanning Electron Microscopy (SEM) revealed changes in the surface morphology of native and treated PHA films. Further, changes in molecular vibrations were confirmed by Fourier Transform Infrared Spectroscopy. [ABSTRACT FROM AUTHOR]

Objective Despite evidence-based guidelines stating that lipase alone should be used in the diagnosis of suspected acute pancreatitis, health care providers continue to order amylase or amylase and lipase together. The purpose of this study was to assess the utilization of appropriate laboratory testing related to the diagnosis of acute pancreatitis. Methods The study used a retrospective cross-sectional design. The timeframe was from January 1, 2020, to December 31, 2020. A retrospective chart review was used to collect data for the following: patient-provider encounter notes, patient demographics, provider demographics, differential and final diagnosis, and laboratory test results. Data analysis include stratification of categorical variables and calculation of cost savings. Results For the 12-month period, this study found 2567 (9.3%) of all amylase and lipase tests to be unnecessary. Amylase tests (1881; 73.2%) made up the most unnecessary tests followed by lipase tests (686; 26.7%). An analysis of test-ordering behavior by providers revealed that 81.5% of all unnecessary tests were ordered by MDs. Finally, this study estimated a total cost savings of $128,350 if all unnecessary tests were eliminated. Conclusion Our study demonstrated that amylase and lipase tests have been overutilized in the diagnosis of acute pancreatitis. [ABSTRACT FROM AUTHOR]

Endophytic bacteria have not been reported widely for their lipolytic abilities, so they are not used for large‐scale lipase production. The purpose of this study was to explore an endophytic bacterium for the production of lipase implementing cost‐effective techniques, including the use of Cassia fistula seeds, as a substrate in the production medium and stem pieces of Phragmites karka as the immobilization matrix. The endophytic strain Bacillus sp. E4 was originally isolated from the halophytic plant Arthrocnemum macrostachyum. Bacillus sp. E4 produced 6.05 IU mL−1 lipase in the presence of powdered seeds of Cassia fistula (golden shower tree). Initial trial experiments using a one‐factor‐at‐a‐time approach led to an improvement in lipase titers to 10.05 IU mL−1. Consequently, investigations using the Plackett–Burman design suggested the influence of three significant factors – incubation period, inoculum size, and substrate concentration – on lipase production. They were optimized using the Box–Behnken design (BBD). In the response optimization experiment, strain E4 yielded 52.35 IU mL−1 lipase, which was in accordance with the predicted yield and indicated an overall 8.65 fold improvement in lipase production. To investigate the use of free cells, strain E4 was immobilized on the stem pieces of a halophytic plant, Phragmites karka, which was used for the first time as an immobilization matrix. The immobilized cells retained lipase production ability for up to six cycles with the highest yield of 110 IU mL−1, which corresponded to an improvement of more than eighteenfold. Scanning electron micrographs confirmed the colonization of E4 cells in the matrix and demonstrated the utilization of C. fistula seeds. Fourier transform infrared spectroscopy affirmed the utilization of components including fatty acids by the immobilized E4 cells. The study suggests that endophytic bacterial strains could be applied for the production of lipase with the utilization of nontraditional oil sources. [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]

The present study assessed the growth and fish performance of juvenile red sea bream fed four isonitrogenous (46–48%) and isolipidic (15%) diets containing 45% fishmeal (control) and 15%, 30%, and 45% defatted black soldier fly (BSF) Hermetia illucens meal for 67 days. Subsequent analyses showed that there were no significant differences in the growth parameters, nutrient digestibility, and body composition of fish among all treatments. Although no significant difference was observed in fish growth, low growth was observed when the fish meal was totally replaced with the BSF meal. Higher alpha diversity of intestinal microbiota was observed in fish fed the BSF meal-based diet compared to that of the control. In the treatment groups fed the BSF meal, elevations in the presence of microbial genera that produce digestive enzymes, such as chitinase (Nocardia, Vibrio, Paenibacillus, Photobacterium), lipase (Paenibacillus, Photobacterium), and trypsin-like serine protease (Bacillus, Paenibacillus) and vitamin B group [biotin (Vibrio, Paenibacillus, Bacillus), cobalamin (Pseudomonas, Bacillus, Propionibacterium), and pyridoxin (Photobacterium)], were observed. We also found an increasing trend of bacteria producing antioxidative enzymes, such as superoxide dismutase and catalase (Nocardia, Pseudomonas, Photobacterium) in the BSF groups. These results suggested that the BSF meal could replace 67% of fishmeal without causing a negative impact on growth and lead to beneficial effects on the intestinal microbiota of the red sea bream. [ABSTRACT FROM AUTHOR]

Agricultural waste and trash are a universal issue which requires extensive working to reduce the impact. Improper management of agricultural waste can lead to serious environmental problems such as soil contamination, water pollution and air pollution. Lipases (protein-based catalyst) were produced with industrial and agriculture by products through solid-state fermentation. Among indigenously isolated strains, procured from Mycology and biotechnology research laboratory, Aspergillus niger (MBL-14) exhibited the highest lipase production, i.e., 7.5 ± 0.63a U/ mL. Various cultural conditions were optimized for maximum production of extracellular lipases. Among various basal substrates, rice bran supported the highest yield of 8.67 ± 0.29a U/mL, ammonium nitrate 6.1 ± 0.95a U/ mL, with the use of sodium chloride 6.1 ± 0.95a U/mL, sucrose 8.0 ± 0.87aU/ mL, the beef extract 9.1 ± 0.84a U/ mL. Maximum extracellular lipases production (9.3 ± 0.55a U/ mL) was achieved through fermentation optimization over a 48-h period, and an increased lipases output (9.6 ± 0.34aU/mL) was obtained at a moistening agent's pH level of 8. Incorporation of oil additives revealed that coconut oil yielded the maximum production (11.0 ± 0.91aU/ mL). Finally, the enzyme was subjected to characterization, at pH 9 after a 1-h incubation period, the enzyme activity was highest, and enzyme was stable. After 40 °C (10.6 ± 0.88aU/mL), the enzyme activity starts decreasing and reaches its lowest at 70 °C. When exposed to the cadmium chloride, the crude enzyme exhibited the highest lipase production (10.1 ± 0.76a U/mL and 10.72 ± 0.85a U/mL) before and after a 1 h of incubation, respectively. [ABSTRACT FROM AUTHOR]

SshEstI, a carboxylesterase from the thermoacidophilic archaeon Saccharolobus shibatae , is a member of the hormone-sensitive lipase family that displays slightly alkaliphilic activity with an optimum activity at pH 8.0. In this study, three distinct strategies were explored to confer acidophilic properties to SshEstI. The first strategy involved engineering the oxyanion hole by replacing Gly81 with serine or aspartic acid. The G81S mutant showed optimum activity at pH 7.0, whereas the aspartic acid mutant (G81D) rendered the enzyme slightly acidophilic with optimum activity observed at pH 6.0; however, k cat and k cat / K m values were reduced by these substitutions. The second strategy involved examining the effects of surfactant additives on the pH-activity profiles of SshEstI. The results showed that cetyltrimethylammonium bromide (CTAB) enhanced wild-type enzyme (WT) activity at acidic pH values. In the presence of 0.1 mM CTAB, G81S and G81D were acidophilic enzymes with optimum activity at pH 6.0 and 4.0, respectively, although their enzyme activities were low. The third strategy involved engineering the active site to resemble that of kumamolisin-As (kuma-As), an acidophilic peptidase of the sedolisin family. The catalytic triad of kuma-As was exchanged into SshEstI using site-directed mutagenesis. X-ray crystallographic analysis of the mutants (H274D and H274E) revealed that the potential hydrogen donor–acceptor distances around the active site of WT were fully maintained in these mutants. However, these mutants were inactive at pH 4–8. [ABSTRACT FROM AUTHOR]

Grape seed oil (GSO) is a valuable resource for the pharmaceutical, cosmetic, and food industries due to its fatty acid composition, which provides anti-inflammatory, cardioprotective, antimicrobial, and anticancer properties. GSO is also recognized for its sensory attributes and serves as an essential component in our dietary intake, providing vital fatty acids for metabolic reactions. The enzymatic synthesis of structured lipids (SL) using GSO was investigated in this study. The SL was created by incorporating medium chain fatty acids (capric acid – C10) at the sn -1 and sn -3 positions and long chain fatty acids in the internal position of the triacylglycerols (TAG). GSO was selected based on its composition rich in unsaturated fatty acids, mainly linoleic acid. The acidolysis reactions were catalyzed by an immobilized commercial lipase Lipozyme TLIM®. The incorporation degree of C10 into GSO reached 36%. Health lipid indices were calculated to determine the C10 incorporation effect on the improvement of nutritional quality of oil. The SL showed lower oxidative stability index (OSI) values than the original oil and higher degree of fatty acid (DFA) values. The simpler, solvent-free method not only enhances the health benefits of oils but also gives valuable properties to GSO, making it an even more attractive option for food industries. [Display omitted] • Sustainable green processes for valorization of grape seed are proposed. • Enzymatic synthesis of structured lipids enhances GSO with health promoting C10 fatty acids. • Lipozyme TLIM¬Æ achieved a remarkable 36% C10 incorporation in GSO. • Simpler, solvent-free method improves GSO's health lipid indices, making it an ideal choice for food industries. [ABSTRACT FROM AUTHOR]

Rhizopus oryzae lipase (ROL) is known to present high selectivity in chemical reactions. However, the poor stability of ROL effectively limits its industrial applications. In this study, several immobilization protocols, such as hydrophobic adsorption, covalent immobilization, multi‐point covalent attachment, ionic adsorption/cross‐linking, and ionic interaction, were applied to improve the stability of ROL. Heterogeneous modification of aspartic and glutamic acid residues on the surface of ROL was carried out by 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide (EDC) to introduce new amine groups with lower pKb. The highest immobilization yield of 89% was achieved for octyl‐agarose, producing specific activity of 45 U/mg, which is 15 folds higher than the specific activity of the soluble enzyme. Improved stability of ROL was observed, in particular for those derivatives obtained by multi‐point covalent attachment of ROL on glyoxyl‐agarose (Gx‐ROL) and aminated ROL on glyoxyl‐agarose (Gx‐NH2‐ROL) by retaining 28%–36% of their initial activities after 24 h incubation at 60°C. Immobilization also altered the co‐solvent stability profile of the immobilized derivatives producing biocatalysts with varied co‐solvent stabilities. Furthermore, utilization of the immobilized preparations in fish oil hydrolysis revealed the selective release of cis‐5,8,11,14,17‐eicosapentaenoic acid (EPA) and cis‐4,7,10,13,16,19‐docosahexaenoic acid (DHA) in favor of EPA. The highest EPA/DHA selectivity of 33 was observed for the hydrophobically immobilized ROL on octyl‐sepharose. [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]

Acute pancreatitis (AP) is a common gastrointestinal disease with high morbidity and mortality rate. Unfortunately, neither the etiology nor the pathophysiology of AP are fully understood and causal treatment options are not available. Recently we demonstrated that heparanase (Hpa) is adversely involved in the pathogenesis of AP and inhibition of this enzyme ameliorates the manifestation of the disease. Moreover, a pioneer study demonstrated that Aspirin has partial inhibitory effect on Hpa. Another compound, which possesses a mild pancreato‐protective effect against AP, is Trehalose, a common disaccharide. We hypothesized that combination of Aspirin, Trehalose, PG545 (Pixatimod) and SST0001 (Roneparstat), specific inhibitors of Hpa, may exert pancreato‐protective effect better than each drug alone. Thus, the current study examines the pancreato‐protective effects of Aspirin, Trehalose, PG545 and SST0001 in experimental model of AP induced by cerulein in wild‐type (WT) and Hpa over‐expressing (Hpa‐Tg) mice. Cerulein‐induced AP in WT mice was associated with significant rises in the serum levels of lipase (X4) and amylase (X3) with enhancement of pancreatic edema index, inflammatory response, and autophagy. Responses to cerulein were all more profound in Hpa‐Tg mice versus WT mice, evident by X7 and X5 folds increase in lipase and amylase levels, respectively. Treatment with Aspirin or Trehalose alone and even more so in combination with PG545 or SST0001 were highly effective, restoring the serum level of lipase back to the basal level. Importantly, a novel newly synthesized compound termed Aspirlose effectively ameliorated the pathogenesis of AP as a single agent. Collectively, the results strongly indicate that targeting Hpa by using anti‐Hpa drug combinations constitute a novel therapy for this common orphan disease. [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 one‐pot stereoselective synthesis of (S)‐flavanones from 2′‐hydroxyacetophenone and substituted aromatic aldehydes was obtained by a double‐face promiscuous chemo‐enzymatic cascade of porcine pancreas and Mucor javanicus lipases. The reaction pathway comprises: A) cross‐aldol condensation catalysed by porcine pancreas lipase to yield chalcone intermediates; B) unprecedented intramolecular oxa‐Michael addition of chalcone intermediates to (S)‐flavanones. Mucor javanicus lipase was the most effective enzyme in step B. Imidazole and 2‐methylimidazole were studied as additive in order to improve the efficacy of the overall transformation. The sustainability of the chemo‐enzymatic cascade was increased by immobilization of lipases on cross‐linked hydroxy‐methylated kraft lignin nanoparticles, by use of concanavalin A. Immobilization conferred considerable stability and reusability at the system for 4 runs. Noteworthy, the reaction mixture was significantly enriched in (S)‐flavanones under both homogeneous and heterogeneous conditions. Computational studies encompassing docking and molecular dynamic analyses showed the role played by evolutionary conserved oxyanion holes and catalytic triad of Mucor javanicus lipase in the stereocontrol of the intra‐molecular oxa‐Michael addition. [ABSTRACT FROM AUTHOR]

Deep eutectic solvents (DESs) are gaining rapid prominence in numerous biocatalysis processes due to their green characteristics, biodegradability, low cost, and simple preparation compared to harsh organic solvents and toxic ionic liquids. However, natural enzymes tend to show activity reduction, even inactivation in the presence of many DESs. Here, we present the first rational design approach to achieve enzymes resistant to both DESs and high temperatures. Using the interaction pattern between DESs and BSLA (Bacillus subtilis lipase A, our model enzyme) derived from all-atom molecular dynamics (MD) simulations, we formulated a solvation-guided engineering strategy. This was established after assessing 33 structural, solvation, and energy observables. We rationally designed and experimentally tested 36 single substitutions, of which 28 (a 77.78% success rate) exhibited improvements in at least two DES cosolvents: choline chloride (ChCl) : acetamide, tetrabutylphosphonium bromide (TBPB) : ethylene glycol (EG), and ChCl : EG. Additionally, through stepwise recombination, we identified two robust BSLA variants, D64H/R107L/E171Y and D64H/R142L, showing stability improvements of up to 4.4-fold and 3.2-fold in three DESs and at 50 °C, respectively. Further MD studies demonstrated that (i) the restricted overall structure but fine-tuned local flexibility and (ii) increased water but decreased DES molecules at substituted sites are two main factors contributing to the enhanced multiple DES resistance. Overall, solvation-guided engineering offers an efficient and rational approach for designing lipases tolerant to DES cosolvents and elevated temperatures, with a considerable potential for adaptation to other enzymes. [ABSTRACT FROM AUTHOR]

Cold‐adapted, alkali‐stable lipases with surfactant tolerance are highly sought after in the detergent industry. In this study, we recombinantly expressed and characterized a novel lipase from Marinobacter nanhaiticus (MNL). The purified MNL exhibited high stability within a temperature range of 10–30°C and pH values of 8–10, demonstrating optimal activity at 20°C and pH 8. MNL has a preference for substrates with medium chains with the best activity being 3768.6 U/mg using p‐nitrophenyl decanoate as a substrate. Remarkably, MNL showed enhanced enzymatic activity in the presence of ionic surfactants and displayed notable resilience when exposed to proteases. Washing performance analysis further revealed MNL's high proficiency in removing oil‐based stains from fabrics. Collectively, these results suggest that MNL holds significant promise as a valuable component in laundry detergent formulations. [ABSTRACT FROM AUTHOR]

Rapeseed oil is a widely consumed edible oil that contains varieties of beneficial micronutrients such as tocopherols and phytosterols; however, the high acid value due to increased free fatty acid can imperil the oil quality and safety. This paper proposed the enzymatic deacidification for high-acid rapeseed oil and simultaneous production of functional diacylglycerols (DAGs) catalyzed by self-made immobilized lipase CALB@MCM-41-C8. The results indicate that the carrier of molecular sieve MCM-41 exhibited a sufficient surface area of 1439.9 m2/g and a proper pore size of 3.5 nm, promoting the immobilization of lipase CLAB. Under the optimal reaction conditions, the acid value of rapeseed oil was largely decreased from 15.3 mg KOH/g to 1.7 mg KOH/g within 3 h, while DAG content was increased from 1.2% to 40.2%. The antioxidant stability of rapeseed oil was also increased from 4.3 h to 7.6 h after enzymatic deacidification. Besides, the deacidified rapeseed oil exhibited fatty, bitter almond aromas, compared to the picked-vegetable, spicy, and pungent aromas for high-acid oil. Finally, the catalytic stability and applicability of CALB@MCM-41-C8 was validated, thus demonstrating the great potential of CALB@MCM-41-C8 in green refining of edible oils and sustainable synthesis of functional lipids. [ABSTRACT FROM AUTHOR]