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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]

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]

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]

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]

Background: In this research, a brewer's yeast suspension was used to biotreat raw linen fibers under a range of different circumstances utilizing an ultrasonic cleaner device. In order to optimize circumstances for the treatment process, this extensive work is focused on examining the variables that could affect the biotreatment, such as the amount of brewer's yeast used, the duration, the temperature of the treatment, and the pH throughout the treatment. After enzymatic treatment, the printing process utilizing turmeric natural dye was used. Variable assesses were conducted to determine the steaming time, thermofixation time, pH of the printing paste, types of dyes, and types of fabrics. How these elements affected the wettability and fabric color strength is investigated. To better comprehend, scanning electron microscope (SEM) was used to study the morphology of treated and untreated linen samples. The effects of treating the fibers with yeast enzyme on their multifunctional qualities, such as color and antibacterial activity against gram-positive bacteria like Staphylococcusaureus and gram-negative bacteria like Escherichia coli, were assessed. Results: Results demonstrated that the enzyme extract, which predominantly contains lipase, amylase, and protease enzymes that develop the fabric printability, is responsible for the increase of color strength which increased by about 152.27% with good fastness properties compared by the untreated printed samples. Conclusions: The overall findings showed that the treated fabrics have superior color fastness and antibacterial properties when compared to the untreated fabrics, demonstrating that the procedure of production used to create these multifunctional linen fabrics is environmentally friendly. [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]

Vicinal halohydrins are key building blocks to produce bioactive molecules and drugs, especially if they can be obtained in enantiomerically pure form. In this study, we present a bi‐enzymatic sequence that allows to obtain vic‐halohydrins through a photochemoenzymatic olefin hydroxy halogenation followed by a lipase catalysed kinetic resolution. The absolute configuration of the resulting products was determined using Mosher's method [ABSTRACT FROM AUTHOR]

Background: Family partial lipodystrophy (FPLD) is a rare autosomal dominant disease characterized by disorders of variable body fat loss associated with metabolic complications. FPLD6 has only been reported in a limited number of cases. Here, we reported a Chinese FPLD6 patient with compound heterozygous mutations in the lipase E, hormone-sensitive type (LIPE) gene. Case presentation: A 20-year-old female patient presented with hypertriglyceridemia, diabetes mellitus, hepatomegaly, and hepatic steatosis. Subcutaneous fat was significantly diminished in her face, abdomen, and limbs. The patient was assessed by detailed clinical and biochemical examinations. A liver biopsy showed severe lipodystrophy. In addition, there were retinal changes, peripheral nerve damage, and renal tubular injury. Sequencing was performed on extracted DNA. Genetic analysis revealed that the patient had compound heterozygous mutations in the LIPE gene: c.2497_ 250ldel (p.Glu833LysfsTer22) and c.2705del (p.Ser902ThrfsTer27) heterozygous mutations. Verification revealed that this mutation was inherited from her father and mother, respectively, and that they formed newly discovered compound heterozygous mutations occurring in the LIPE gene, causing FPLD6. Conclusion: We reported the first case of FPLD6 in China. Gene analysis demonstrated compound heterozygous mutations in LIPE in this patient. Our case emphasizes the importance of genetic testing in young patients with severe metabolic syndromes. [ABSTRACT FROM AUTHOR]

Objective: Several studies have investigated hormones such as growth hormone (GH), insulin (INS), insulin-like growth factor-1 (IGF-1), and hormone-sensitive lipase (HSL). However, there is insufficient data on the effects of the combination of short-, medium-, and long-chain fatty acids on hormone concentrations in serum/tissue, including GH, INS, IGF-1, and HSL. The purpose of this study was to investigate the effects of butyric acid (BA), caprylic acid (CA), and oleic acid (OA) alone or in combination on GH, INS, IGF-1, and HSL secretion. Methods: Fifty-six male Wistar rats were used in the study. The animals were separated into 8 subgroups: control, BA, CA, OA, BA + CA, BA + OA, CA + OA, and BA + CA + OA groups. Fatty acids were administered orally to rats for 21 days. At the end of the study, GH, IGF, INS, and HSL levels were measured in serum using the enzyme-linked immunosorbent assay method. Results: BA administration reduced GH, IGF-1, and INS levels but had no significant effect on HSL levels. CA administration increased HSL levels but had no significant effect on GH, INS, and IGF-1 levels. OA administration increased GH and HSL levels but had no significant effect on IGF-1 and INS levels. Conclusion: The combined use of fatty acids increased GH levels while decreasing INS, IGF-1, and HSL levels. [ABSTRACT FROM AUTHOR]

The present research examined the impact of L-glutamic acid (Glu) supplementation on the growth performance, muscle composition, gene expression correlated with muscle growth, and intestinal health of largemouth bass. There were 525 fish in total, which were distributed randomly into five groups. Each group had three replicates, and each replicate consisted of 35 fish. Groups with control and experimental diets were assigned glutamic acid amounts of 0.2%, 0.4%, 0.6%, and 0.8%. The findings demonstrated that glutamic acid supplementation enhanced growth performance, feed intake (FI), and condition factor (CF), with the best value being attained at 0.4% Glu. The mean muscle fiber area was increased and the muscle fiber density was decreased in the 0.6% Glu group. The levels of total amino acids and specific amino acids, such as glutamic acid, aspartic acid, leucine, valine, alanine, and glycine, were shown to be higher in the 0.6% Glu group. In the 0.6% Glu group, the mRNA expression levels of atrogin-1, murf-1, foxo3a, and 4e-bp1 were decreased compared to the control group. Conversely, the mRNA expression levels of myf5, myog, myod, s6k1, tor, akt, and pi3k were increased in the 0.6% Glu group compared to the control group. The 0.4% Glu group had higher intestinal amylase, lipase, and protease activities and greater villus height, villus width, and muscle thickness. In summary, Glu can support largemouth bass growth, muscular development, intestinal digestion, and absorption. [ABSTRACT FROM AUTHOR]

In this study, the wild-type Bacillus cereus ATA179 was mutagenized by random UV mutagenesis to increase lipase production. The mutant with maximum lipolytic activity was named Bacillus cereus EV4. The mutant strain (10.6 U/mL at 24 h) produced 60% more enzyme than the wild strain (6.6 U/mL at 48 h). Nutritional factors on lipase production were investigated. Sucrose was the best carbon source, (NH4)2HPO4 was the best nitrogen source and CuSO4 was the best metal ion source. Mutant EV4 showed a 32% increase in lipase production in the modified medium. The optimum temperature and pH were found to be 60 °C and 7.0, respectively. CuSO4, CaCl2, LiSO4, KCl, BaCl2, and Tween 20 had an activating effect on the enzyme. Vmax and Km values were found to be 17.36 U/mL and 0.036 mM, respectively. The molecular weight was determined as 28.2 kDa. The activity of lipase was found to be stable up to 60 days at 20 °C, 75 days at 4 °C, and 90 days at −20 °C. The potential of lipase in the detergent industry was investigated. The enzyme was not affected by detergent additives but was effective in removing stains in fabrics contaminated with oily substances. [ABSTRACT FROM AUTHOR]

Summary: Processing by proteases irreversibly regulates the fate of plant proteins and hampers the production of recombinant proteins in plants, yet only few processing events have been described in agroinfiltrated Nicotiana benthamiana, which has emerged as the main transient protein expression platform in plant science and molecular pharming.Here, we used in‐gel digests and mass spectrometry to monitor the migration and topography of 5040 plant proteins within a protein gel. By plotting the peptides over the gel slices, we generated peptographs that reveal where which part of each protein was detected within the protein gel.These data uncovered that 60% of the detected proteins have proteoforms that migrate at lower than predicted molecular weights, implicating extensive proteolytic processing. This analysis confirms the proteolytic removal and degradation of autoinhibitory prodomains of most but not all proteases, and revealed differential processing within pectinemethylesterase and lipase families. This analysis also uncovered intricate processing of glycosidases and uncovered that ectodomain shedding might be common for a diverse range of receptor‐like kinases. Transient expression of double‐tagged candidate proteins confirmed processing events in vivo.This large proteomic dataset implicates an elaborate proteolytic machinery shaping the proteome of N. benthamiana. [ABSTRACT FROM AUTHOR]

Microflora of reptiles and amphibians pose a serious threat to the environment and humans, as these bacteria are more resistant to antimicrobials or have more virulence factors. In this study, we focused on the characterization of Gram-negative bacterial isolates obtained from Mauremys rivulata (Western Caspian turtle), Natrix natrix (Grass snake), Natrix tessellata (Dice snake) and Pelophylax bedriagae (Levantine frog) using cultural methods and determine some virulence factors (antibiotic resistance, VanA and TetM resistant genes, biofilm, siderophore and some enzymatic activities etc.). Multiple antibiotic resistance (MAR) and virulence index (VI) of each strain were also calculated. A total of 85 Gram-negative isolates exhibited varying levels of antibiotic resistance and the ability to produce one or more extracellular enzymes such as amylase, lecithinase, protease, lipase, hemolysis, siderophore, amino acid decarboxylase and DNase. Although the TetM has been identified in all 7 isolates, none of the isolates have shown the presence of the VanA gene. According to MAR and VI results, most of the isolates were found to be in the high threat group and these indexes were correlated with antibiotic resistance. The study data revealed that chemicals used in industry and agriculture both trigger environmental pollution and pose risk factors for wildlife. [ABSTRACT FROM AUTHOR]

Background: Prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) is higher in men than in women. Hormonal and genetic causes may account for the sex differences in MASLD. Current human in vitro liver models do not sufficiently take the influence of biological sex and sex hormones into consideration. Methods: Primary human hepatocytes (PHHs) were isolated from liver specimen of female and male donors and cultured with sex hormones (17β-estradiol, testosterone and progesterone) for up to 72 h. mRNA expression levels of 8 hepatic lipid metabolism genes were analyzed by RT-qPCR. Sex hormones and their metabolites were determined in cell culture supernatants by LC-MS analyses. Results: A sex-specific expression was observed for LDLR (low density lipoprotein receptor) with higher mRNA levels in male than female PHHs. All three sex hormones were metabolized by PHHs and the effects of hormones on gene expression levels varied depending on hepatocyte sex. Only in female PHHs, 17β-estradiol treatment affected expression levels of PPARA (peroxisome proliferator-activated receptor alpha), LIPC (hepatic lipase) and APOL2 (apolipoprotein L2). Further changes in mRNA levels of female PHHs were observed for ABCA1 (ATP-binding cassette, sub-family A, member 1) after testosterone and for ABCA1, APOA5 (apolipoprotein A-V) and PPARA after progesterone treatment. Only the male PHHs showed changing mRNA levels for LDLR after 17β-estradiol and for APOA5 after testosterone treatment. Conclusions: Male and female PHHs showed differences in their expression levels of hepatic lipid metabolism genes and their responsiveness towards sex hormones. Thus, cellular sex should be considered, especially when investigating the pathophysiological mechanisms of MASLD. [ABSTRACT FROM AUTHOR]

Noncovalent interactions associated with the enzyme, enzyme-substrate complexes, and transition states have profound influences on varying the biocatalytical activity and enantioselectivity. The recent rediscovery of halogen bonds incorporating in biocatalysis has induced our motivation on introducing halogen substituents into the substrate for improving the enzyme performance. By selecting Candida antarctica lipase B (CALB)-catalyzed kinetic hydrolysis (or butanolysis) of trans-2-(p-halophenyl)cyclopropyl 1,2,4-triazolide in methyl tert-butyl ether (MTBE) as the model reaction system, (R, R)-2-(p-halophenyl)cyclopropane-1-carboxylic acid (or ester) of high-optical purity that acts as the key building block for synthesizing the corresponding (1 R, 2S)-2-(p-halophenyl)cyclopropylamine derivatives was obtainable. The thermodynamic and kinetic analysis was then employed for elucidating effects of the halogen substituent and acyl acceptor on enhancing the noncovalent interactions of the rate-limiting tetrahedral intermediate and hence CALB activity and enantioselectivity. A putative model was moreover proposed for interpreting the possible hydrogen and halogen bonding involving the halogen substituent in turning the noncovalent interactions. [ABSTRACT FROM AUTHOR]

The culturable yeast communities in temperate forest soils under the ornithogenic influence were studied in a seasonal dynamic. To investigate the intense ornithogenic influence, conventional and "live" feeders were used, which were attached to trees in the forest and constantly replenished throughout the year. It was found that the yeast abundance in the soil under strong ornithogenic influence reached the highest values in winter compared to the other seasons and amounted to 4.8 lg (cfu/g). This was almost an order of magnitude higher than the minimum value of yeast abundance in ornithogenic soils determined for summer. A total of 44 yeast species, 21 ascomycetes and 23 basidiomycetes, were detected in ornithogenic soil samples during the year. These included soil-related species (Barnettozyma californica, Cyberlindnera misumaiensis, Cutaneotrichosporon moniliiforme, Goffeauzyma gastrica, Holtermanniella festucosa, Leucosporidium creatinivorum, L. yakuticum, Naganishia adeliensis, N. albidosimilis, N. globosa, Tausonia pullulans, and Vanrija albida), eurybionts (yeast-like fungus Aureobasidium pullulans, Debaryomyces hansenii, and Rhodotorula mucilaginosa), inhabitants of plant substrates and litter (Cystofilobasidium capitatum, Cys. infirmominiatum, Cys. macerans, Filobasidium magnum, Hanseniaspora uvarum, Metschnikowia pulcherrima, and Rh. babjevae) as well as a group of pathogenic and opportunistic yeast species (Arxiozyma bovina, Candida albicans, C. parapsilosis, C. tropicalis, Clavispora lusitaniae, and Nakaseomyces glabratus). Under an ornithogenic influence, the diversity of soil yeasts was higher compared to the control, confirming the uneven distribution of yeasts in temperate forest soils and their dependence on natural hosts and vectors. Interestingly, the absolute dominant species in ornithogenic soils in winter (when the topsoil temperature was below zero) was the basidiomycetous psychrotolerant yeast T. pullulans. It is regularly observed in various soils in different geographical regions. Screening of the hydrolytic activity of 50 strains of this species at different temperatures (2, 4, 10, 15 and 20 °C) showed that the activity of esterases, lipases and proteases was significantly higher at the cultivation temperature. Ornithogenic soils could be a source for the relatively easy isolation of a large number of strains of the psychrotolerant yeast T. pullulans to test, study and optimize their potential for the production of cold-adapted enzymes for industry. [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]

The low cost and excellent catalytic properties of lipase for industrial processes are highly desirable. A promising new approach involves the support modification of lipase and spacer arm, which enables the enhancement of lipase properties. This study investigates the immobilization of crude lipase from Mucor miehei onto a Polyurethane Foam (PUF) surface using various coating techniques. The PUF matrix was obtained through isocyanate and polyol reactions. Subsequently, the PUF was coated by adsorbing lipase and adding edible support material. The immobilized lipase was then utilized in the hydrolysis of coconut oil to produce fatty acids. Furthermore, the immobilized enzyme was employed in the esterification of fatty acids to produce bio-flavors. The results demonstrate that the attachment reaction using support material, namely lecithin, gelatin, MgCl2, and Polyethylene glycol 6000 (PEG), all of which are simple and edible, was able to enhance the stability and reusability of lipase. This immobilization technique increased triglyceride hydrolysis into FFA by 422%. The successful edible support modification of immobilized lipase from M. miehei on PUF, coupled with significantly enhanced enzyme stability and catalytic activity, offers a promising, environmentally friendly solution for diverse applications in the food industry. [ABSTRACT FROM AUTHOR]

Background: Cutibacterium (C. acnes) lipase activity can be reduced by topical antifungal cream called ketoconazole (KTZ). Additionally, it contains anti-inflammatory and anti-androgenic properties. The aim of this study was to evaluate the efficacy of topical ketoconazole 2% cream versus placebo in the treatment of mild and moderate acne vulgaris. Patients & Methods: 40 patients presented by mild and moderate acne were instructed to apply ketoconazole 2 % cream to the right half of the face and placebo to the left half of the face twice daily for 8 weeks Results: The mean percent reduction of inflammatory lesions (papules and pustules) and Total lesions count from baseline was significantly higher in ketoconazole than placebo. There was a statistically significant decrease in acne severity assessed by IGA score in both sides. The success rate of ketoconazole was significantly higher than placebo. In ketoconazole side, 17.5% of patients considered clear versus12.5%in control side and 52.5% almost clear versus 35% in control side. There was a statistically significant negative relationship between the therapeutic response and number of pustules in each side. Conclusion: Topical ketoconazole 2% can be an effective and safe treatment option for mild and moderate acne vulgaris. [ABSTRACT FROM AUTHOR]

Entrapment is a strategy widely used for lipase immobilization and recovery. However, mass transfer issues and the high costs of bead production limit the potential of the technique. This study aimed to investigate the impacts associated with the replacement of sodium alginate with corn starch on bead formation by jet cutting. BET analysis showed surface areas of 1.02 and 1.24 m2 g−1 for beads containing 25 % (B25) and 50 % (B50) starch, respectively. XPS showed nitrogen distribution rates of 7.40–8.77 % (B25) and 4.75–5.10 % (B50). Both immobilized derivatives achieved activity recoveries of 56.23 % ± 5.73 % (B25) and 51.58 % ± 5.56 % (B50) and maximum specific activity of 1500 U g−1 protein. The thermal inactivation constant values of free lipase were 0.0096 and 0.0285 min−1 at 70 and 75 °C, while to immobilized lipases were 7-fold and 2-fold lower than that at 70 and 75 °C, respectively. The half-life time of free lipase and immobilized derivatives (B25 and B50), at 70º C, were 72.20, 577.62 and 533.19 min, respectively. Biodiesel yields of 50.49 % ± 5.67 % and 51.16 % ± 9.91 % were achieved using B25 and B50, respectively. These findings demonstrate lipase can be successfully immobilized on alginate–starch beads by jet cutting. [Display omitted] • Alginate–starch beads were produced by jet cutting. • Eversa Transform 2.0 was successfully entrapped in alginate–starch beads. • XPS showed a uniform nitrogen distribution in inner and outer layers. • The k d of free lipase was 7.38- and ∼2-fold higher than that of immobilized lipases. • Both immobilized lipases were able to produce biodiesel. [ABSTRACT FROM AUTHOR]

[Display omitted] The catalytic performance of immobilized lipase is greatly influenced by functional support, which attracts growing interest for designing supports to achieve their promotive catalytic activity. Many lipases bind strongly to hydrophobic surfaces where they undergo interfacial activation. Herein, the behavioral differences of lipases with distinct lid structures on interfaces of varying hydrophobicity levels were firstly investigated by molecular simulations. It was found that a reasonable hydrophilic/hydrophobic surface could facilitate the lipase to undergo interfacial activation. Building on these findings, a novel "nest"-like superhydrophobic ZIFs (ZIFN) composed of hydrophobic ligands was prepared for the first time and used to immobilize lipase from Aspergillus oryzae (AOL@ZIFN). The AOL@ZIFN exhibited 2.0-folds higher activity than free lipase in the hydrolysis of p-Nitrophenyl palmitate (p-NPP). Especially, the modification of superhydrophobic ZIFN with an appropriate amount of hydrophilic tannic acid can significantly improve the activity of the immobilized lipase (AOL@ZIFN-TA). The AOL@ZIFN-TA exhibited 30-folds higher activity than free lipase, and still maintained 82% of its initial activity after 5 consecutive cycles, indicating good reusability. These results demonstrated that nanomaterials with rational arrangement of the hydrophilic/hydrophobic surface could facilitate the lipase to undergo interfacial activation and improve its activity, displaying the potential of the extensive application. [ABSTRACT FROM AUTHOR]

Docosahexaenoic acid plays a crucial role in infant brain function, and the market demand of high-purity docosahexaenoic acid is continuously increasing. The availability of docosahexaenoic acid in natural fish oil is limited, prompting the exploration of alternative sources like microalgae. For algal oil, enzymatic ethanolysis is preferred to chemical methods because the former is milder and can avoid docosahexaenoic acid oxidation. However, enzymatic methods have generally low yield due to the poor substrate-specificity of lipase to long-chain polyunsaturated fatty acids, affecting the yield and purity of docosahexaenoic acid. Therefore, we developed an efficient process to produce high-purity docosahexaenoic acid ethyl ester from algal oil, by screening lipases, optimizing enzymatic ethanolysis and applying molecular distillation. Lipase UM1 was the best lipase to produce ethyl ester from algal oil with the highest ethyl ester yield (95.41%). Meanwhile, it was a catalyst for the reaction of long-chain polyunsaturated fatty acids with ethanol. The fatty acid docosahexaenoic acid conversion rates exceeded 90%. After molecular distillation, a final product containing 96.52% ethyl ester was obtained with a docosahexaenoic acid content up to 80.11%. Our findings provide an highly effective enzymatic method for the production of high-purity docosahexaenoic acid ethyl esters, with potential commercial applications. [ABSTRACT FROM AUTHOR]

In recent years, there has been a growing interest in regulating lipid digestion through the construction of various interfacial structures. In the present work, a series of complex interfacial structures were designed by combining Tween 80 in the aqueous phase and lecithin in the oil phase at different concentration ratios. The emulsification properties, the roles in regulating lipid digestion, and the interfacial dilatational rheological properties of the composite emulsifying systems were characterized. The results showed that the combination of Tween 80 and lecithin at different ratios could effectively modulate the rate of lipid digestion. The polyoxyethylene chains of Tween 80 formed a network, that provided a spatial obstacle for the adsorption of bile salts and lipases. Thus, Tween 80 significantly delayed the lipid digestion. The introduction of lecithin gradually replaced Tween 80 molecules at the interface, thus providing space for the adsorption of bile salts and lipases. In addition, as the ratio of lecithin concentration to Tween 80 increased, lecithin gradually became the dominant factor in the interfacial properties. As a result, the rate of lipid digestion was accelerated. Therefore, by compounding different ratios of lecithin and Tween 80, a series of emulsions with different lipid digestion rates were obtained. This research provides a basis for rationally designing food emulsions according to specific needs. [ABSTRACT FROM AUTHOR]

This study developed three microstructured chitosan nanocapsules with immobilized lipase to explore chitosan-lipase interactions at different pH levels. Chitosan undergoes complete protonation or deprotonation based on pH level. Three distinct pH levels were examined: 5.5, where chitosan is fully protonated; 6.5, where chitosan is partially protonated/deprotonated; and 7.5, where chitosan is fully deprotonated. The nanocapsules exhibited nanoscale dimensions and the microstructures showed porous morphology. Immobilized lipase showed improved temperature stability, compared to free enzyme, especially in lipase supports at pH 5.5 and 7.5 due to electrostatic and hydrophobic interactions. The interactions between chitosan and lipase influenced the microenvironment around the active site, resulting in an optimum pH of 8 for all supports. Immobilized lipase at pH 5.5 and 7.5 displayed the best reusability in the hydrolysis of p-nitrophenyl palmitate under reaction conditions of 37 °C and pH 8. During refrigeration storage, all immobilized lipases maintained total activity for 7 days, but lipase immobilized at pH 6.5 maintained more the activity after 28 days. Therefore, this study has developed promising immobilized lipase, standing out not only for industrial application concerning cost-effectiveness, but also for the innovation in investigating the influence of chitosan-lipase interactions during immobilization. [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]

The rising prevalence of diabetes and the high cost of treatment necessitate the search for natural, safe, and effective treatment methods. This study examined the in vitro and in silico pancreatic lipase inhibitory activity as well as the hypoglycemic effect of the ethanol extract of Tapinanthus cordifolius (TC) leaves in mice. TC leaves were extracted with ethanol by Soxhlet extraction technique. The pancreatic lipase inhibitory activity of TC extract at various concentrations (50, 100, 200, 400, 800, and 1600 µg/mL) was evaluated using spectrophotometric method. The effect of bioactive compounds from TC extract on pancreatic lipase was assessed in silico using molecular docking approach. The hypoglycaemic effect of TC extract was evaluated in vivo by an oral glucose tolerance test (OGTT) in normoglycaemic mice. Glibenclamide (5 mg/kg) was used as the positive control. TC extract showed significant pancreatic lipase (PL) inhibitory activity with IC50 value of 294.30 ± 0.67 µg/mL, which was comparable to that of orlistat (IC50 = 195.70 ± 4.60 µg/mL). In silico studies identified campesterol, heptadecafluorononanoic acid, α-tocopherol-β-d-mannoside, clionasterol, and β-tocopherol as potent pancreatic lipase inhibitors. TC extract (200, 400, and 800 mg/kg) significantly reduced hyperglycemia in glucose-challenged mice. It exhibited a significant (p<0.05) increase in glucose absorption, similar to glibenclamide, with the most notable effect at 60 minutes post-glucose load. These findings suggest that Tapinanthus cordifolius leaf can effectively inhibit pancreatic lipase and reduce blood glucose levels, and may serve as a natural treatment alternative for diabetes mellitus. [ABSTRACT FROM AUTHOR]

This study presents a novel approach to producing activated carbon from agro-industrial residues, specifically cocoa fruit peel, using solid-state fermentation (SSF) with Aspergillus niger. The process effectively degrades lignin, a major impediment in traditional activated carbon production, resulting in a high-quality carbon material. This carbon was successfully utilized for enzyme immobilization and aroma synthesis, showcasing its potential as a versatile biocatalyst. The study meticulously evaluated the physical and chemical attributes of activated carbon derived from fermented cocoa peel, alongside the immobilized enzymes. Employing a suite of analytical techniques—electrophoresis, FTIR, XRD, and TG/DTG the research revealed that fermentation yields a porous material with an expansive surface area of 1107.87 m2/g. This material proves to be an excellent medium for lipase immobilization. The biocatalyst fashioned from the fermented biomass exhibited a notable increase in protein content (13% w/w), hydrolytic activity (15% w/w), and specific activity (29% w/w), underscoring the efficacy of the fermentation process. The significant outcome of this research is the development of a sustainable method for activated carbon production that not only overcomes the limitations posed by lignin but also enhances enzyme immobilization for industrial applications. The study's findings have important implications for the agro-industrial sector, promoting a circular economy and advancing sustainable biotechnological processes. [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]

Background: Litter size plays a crucial role in determining profitability in the sheep industry. Breeding sheep with high litter sizes could be enhanced by selecting candidate genes. One gene affecting sheep's reproductive performance is the hormone-sensitive lipase (HSL) gene. As a result, this study investigated whether the HSL gene variation influenced the fertility of Awassi ewes. The genomic DNA was extracted from 52 singleton ewes and 48 twin ewes. The HSL gene exon 9 (278 bp) was amplified using polymerase chain reaction (PCR). Results: Study results revealed two genotypes identified in the 278-bp amplicons: GG and GA. Molecular sequence analysis identified a novel mutation in the GA genotype 216G > A. The statistical analysis revealed a significant association between the single nucleotide polymorphism (SNP) 216A > G and reproductive performance. Ewes with the SNP 216G > A genotype exhibited significantly increased litter sizes, twinning rates, lambing rates, and fewer days to lambing compared to ewes with GG genotypes (P ≤ 0.05). The logistic regression analysis results provided strong evidence that the 216G > A mutation significantly increased litter sizes. Conclusions: This study concluded that variant 216G > A SNP positively impacts Awassi sheep reproduction. There is a higher litter size and more prolificacy in ewes with the 216G > A SNP than in those without the SNP. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the effects of S. nigra L. and V. agnuscastus L. plants on obesity in vivo. Extracts were prepared from S. nigra leaves, flowers, fruits and from V. agnus-castus leaves, flowers, and fruits using 100% water and 70% ethanol. The total phenol and flavonoid contents of the extracts were quantified spectrophotometrically. The findings revealed that the ethanol extracts of V. agnus-castus and S. nigra flowers had the highest phenolic content, while the ethanol extracts of S. nigra flowers and V. agnus-castus leaves had the highest flavonoid content. Qualification and quantification of the phenolic contents of the extracts were carried out using liquid chromatography-high resolution mass spectrometry (LC-HRMS) analyses. The study investigated the effects of various extracts on plasma levels of leptin, insulin, triiodothyronine (T3), thyroxine (T4), triglycerides, high-density lipoprotein (HDL), low-density lipoprotein (LDL) and lipase enzyme in obesity-induced rats. The results showed that the ethanol extract of V. agnus-castus flowers, as well as the ethanol and water extracts of V. agnus-castus leaves, resulted in body weight reduction in rats with obesity. Additionally, these extracts were found to decrease serum levels of LDL, triglycerides, leptin, lipase, TNF-α, and IL-1β while increasing levels of HDL and adiponectin. The LC-HRMS results demonstrated that all three extracts exhibited relatively high concentrations of luteolin-7-glycoside and kaempferol, in comparison to the other extracts. The ethanol extract of V. agnus-castus flowers contained 653.04 mg/100 g of luteolin-7-glycoside and 62.63 mg/100 g of kaempferol. The ethanol extract of V. agnus-castus leaves contained 1,720.26 mg/100 g of luteolin-7-glycoside and 95.85 mg/100 g of kaempferol. The water extract of V. agnus-castus leaves contained 690.49 mg/ 100 g of luteolin-7-glycoside and 194.41 mg/100 g of kaempferol. The study suggests that the ethanol extract of V. agnus-castus flowers and leaves, as well as the water extract of V. agnus-castus leaves, may have potential benefits in treating obesity. However, further controlled clinical studies are necessary to evaluate the clinical efficacy of V. agnus-castus in treating obesity and investigate the in vivo anti-obesogenic effects of luteolin-7-glycoside and kaempferol separately, both in their pure form and in combination. [ABSTRACT FROM AUTHOR]

Musa balbisiana Colla plant belongs to the Musaceae family, which is well‐known for its nutritional and pharmacological properties. This study aimed to evaluate the anti‐xanthine oxidase, anti‐lipase, and anti‐tyrosinase activities of samples of corm, pseudostem, inflorescence, fruit, peel, and seed of M. balbisiana. The results show that seed performed the highest capacity in the inhibition of three investigated enzymes, with IC50 values of 290.25 μg/mL (anti‐tyrosinase), 141.51 μg/mL (anti‐xanthine oxidase), and 25.66 μg/mL (anti‐lipase). In addition, while fruit showed better performance in anti‐lipase inhibition (IC50: 32.72 μg/mL), corm pointed out higher capability on anti‐xanthine oxidase (IC50: 160.67 μg/mL) and anti‐tyrosinase (IC50: 334.43 μg/mL). The data show the high potential of the application of M. balbisiana parts in medical aesthetics, dietary supplements, or medicine. This is the first report to systematically present the biological activities of all parts of M. balbisiana grown in Vietnam. [ABSTRACT FROM AUTHOR]

Successful bisexual reproduction requires interactions between males and females. Male‐derived seminal fluid proteins (SFPs) transferred to females during mating profoundly affect females from pre‐ to post‐mating, and the subsequent shift in female physiology enhances their fertility. SFPs have important evolutionary implications for the fitness of many insects. However, little is known about how females respond to male SFPs. In this study, we identified a male‐derived SFP‐phospholipase A2 (PLA2) in Ophraella communa. PLA2 is a vital enzyme in eicosanoid biosynthesis; however, it has not been identified as an insect SFP. We found that OcPLA2 is specifically expressed in males, especially in the male accessory glands (MAGs); it is transferred to the female during mating and functions as an SFP to enhance fertility. The expression of a female‐derived gene encoding the WD repeat‐containing protein 46 (WD46) was upregulated when OcPLA2 entered the female reproductive tract, and this contributed to female egg production by increasing triacylglycerol lipase (TGL) gene expression and the triglyceride (TG) content. This is the first study to identify PLA2 as an SFP in insects. Our findings also shed light on the regulatory role of OcPLA2 in beetle reproduction; the expression of OcPLA2 is initially correlated with female WD46 expression and later with the decline in TGL gene expression and the TG content. This represents a unique mechanism of reproductive regulation by an SFP. [ABSTRACT FROM AUTHOR]

Herein we present the biocatalysed preparation of a mono-N-carbamate-protected precursor of antitumoral Nutlin-3a through enantioselective alkoxycarbonylation of meso-1,2-disubstituted-1,2-diaminoethane using enzyme lipases and dialkyl carbonates as acylating agents. A series of supported or free lipase enzymes were screened in combination with commercially available diallyl, diethyl and dimethyl carbonates. The reactions were conducted at different temperatures, for different reaction times and with variable co-solvent systems to evaluate the effects on the enzyme catalytic activity. The best results in terms of conversion, enantiomeric excess and yield were obtained when lipase from Candida antarctica B (CAL-B) was used with diallyl carbonate (DAC) when conducting the reaction solventless at 75 °C. [ABSTRACT FROM AUTHOR]

Lipases are enzymes that hydrolyze long-chain carboxylic esters, and in the presence of organic solvents, they catalyze organic synthesis reactions. However, the use of solvents in these processes often results in enzyme denaturation, leading to a reduction in enzymatic activity. Consequently, there is significant interest in identifying new lipases that are resistant to denaturing conditions, with extremozymes emerging as promising candidates for this purpose. Lip7, a lipase from Geobacillus sp. ID17, a thermophilic microorganism isolated from Deception Island, Antarctica, was recombinantly expressed in E. coli C41 (DE3) in functional soluble form. Its purification was achieved with 96% purity and 23% yield. Enzymatic characterization revealed Lip7 to be a thermo-alkaline enzyme, reaching a maximum rate of 3350 U mg−1 at 50 °C and pH 11.0, using p-nitrophenyl laurate substrate. Notably, its kinetics displayed a sigmoidal behavior, with a higher kinetic efficiency (kcat/Km) for substrates of 12-carbon atom chain. In terms of thermal stability, Lip7 demonstrates stability up to 60 °C at pH 8.0 and up to 50 °C at pH 11.0. Remarkably, it showed high stability in the presence of organic solvents, and under certain conditions even exhibited enzymatic activation, reaching up to 2.5-fold and 1.35-fold after incubation in 50% v/v ethanol and 70% v/v isopropanol, respectively. Lip7 represents one of the first lipases from the bacterial subfamily I.5 and genus Geobacillus with activity and stability at pH 11.0. Its compatibility with organic solvents makes it a compelling candidate for future research in biocatalysis and various biotechnological applications. [ABSTRACT FROM AUTHOR]

The walnut (Juglans regia L.) is a typical and an economically important tree species for nut production with heterodichogamy. The absence of female and male flowering periods seriously affects both the pollination and fruit setting rates of walnuts, thereby affecting the yield and quality. Therefore, studying the characteristics and processes of flower bud differentiation helps in gaining a deeper understanding of the regularity of the mechanism of heterodichogamy in walnuts. In this study, a total of 3540 proteins were detected in walnut and 885 unique differentially expressed proteins (DEPs) were identified using the isobaric tags for the relative and absolute quantitation (iTRAQ)-labeling method. Among all DEPs, 12 common proteins were detected in all four of the obtained contrasts. GO and KEGG analyses of 12 common DEPs showed that their functions are distributed in the cytoplasm metabolic pathways, photosynthesis, glyoxylate and dicarboxylate metabolism, and the biosynthesis of secondary metabolites, which are involved in energy production and conversion, synthesis, and the breakdown of proteomes. In addition, a function analysis was performed, whereby the DEPs were classified as involved in photosynthesis, morphogenesis, metabolism, or the stress response. A total of eight proteins were identified as associated with the morphogenesis of stamen development, such as stamen-specific protein FIL1-like (XP_018830780.1), putative leucine-rich repeat receptor-like serine/threonine-protein kinase At2g24130 (XP_018822513.1), cytochrome P450 704B1-like isoform X2 (XP_018845266.1), ervatamin-B-like (XP_018824181.1), probable glucan endo-1,3-beta-glucosidase A6 (XP_018844051.1), pathogenesis-related protein 5-like (XP_018835774.1), GDSL esterase/lipase At5g22810-like (XP_018833146.1), and fatty acyl-CoA reductase 2 (XP_018848853.1). Our results predict several crucial proteins and deepen the understanding of the biochemical mechanism that regulates the formation of male and female flower buds in walnuts. [ABSTRACT FROM AUTHOR]