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1.
Lipids Health Dis ; 23(1): 221, 2024 Jul 22.
Article de Anglais | MEDLINE | ID: mdl-39039573

RÉSUMÉ

BACKGROUND: Compared with moderate-intensity continuous training (MICT), high-intensity interval training (HIIT) has at least a comparable effect on inhibiting an increase in fat. However, few studies have been conducted to examine the effects of detraining on body fat in rats fed a high-fat diet. The present study aimed to compare the effects of 10 weeks of HIIT or MICT as well as 6 weeks of detraining on body fat in rats fed a high-fat diet. METHODS: After being fed a high-fat diet for 8 weeks, 54 female rats were randomly assigned to six groups: (1) CON-10, sedentary control for 10 weeks; (2) MICT-10, 10 weeks of MICT; (3) HIIT-10, 10 weeks of HIIT; (4) CON-16, sedentary control for 16 weeks; (5) MICT-16, 10 weeks of MICT followed by 6 weeks of training cessation; and (6) HIIT-16, 10 weeks of HIIT followed by 6 weeks of training cessation. The training was performed 5 days/week. The subcutaneous adipose tissue (inguinal; SCAT), visceral adipose tissue (periuterine; VAT) and serum lipid profile were analysed after 10 or 16 weeks. Adipose tissue triglyceride lipase (ATGL) protein expression in VAT was assessed by western blotting. RESULTS: HIIT-10 and MICT-10 prevented the increase in SCAT, VAT and serum lipid levels seen in the CON group. During the 6-week detraining period, HIIT continued to prevent the increase in adipose tissue mass observed in the CON group, whereas MICT at least maintained this inhibition. The inhibition of fat mass increase was mainly the result of preventing adipocyte hypertrophy. The HIIT-10 and HIIT-16 groups showed the highest ATGL protein expression. CONCLUSIONS: HIIT has a comparable effect to MICT on inhibiting fat accumulation in female rats; however, the inhibition of SCAT and VAT increase by HIIT is superior to MICT after short-term training cessation.


Sujet(s)
Alimentation riche en graisse , Entrainement fractionné de haute intensité , Conditionnement physique d'animal , Animaux , Femelle , Entrainement fractionné de haute intensité/méthodes , Alimentation riche en graisse/effets indésirables , Rats , Graisse intra-abdominale/métabolisme , Triacylglycerol lipase/métabolisme , Rat Sprague-Dawley , Tissu adipeux/métabolisme , Graisse sous-cutanée/métabolisme , Acyltransferases
2.
Mar Drugs ; 22(7)2024 Jun 28.
Article de Anglais | MEDLINE | ID: mdl-39057410

RÉSUMÉ

Omega-3 fatty acids are essential fatty acids that are not synthesised by the human body and have been linked with the prevention of chronic illnesses such as cardiovascular and neurodegenerative diseases. However, the current dietary habits of the majority of the population include lower omega-3 content compared to omega-6, which does not promote good health. To overcome this, pharmaceutical and nutraceutical companies aim to produce omega-3-fortified foods. For this purpose, various approaches have been employed to obtain omega-3 concentrates from sources such as fish and algal oil with higher amounts of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Among these techniques, enzymatic enrichment using lipase enzymes has gained tremendous interest as it is low in capital cost and simple in operation. Microorganism-derived lipases are preferred as they are easily produced due to their higher growth rate, and they hold the ability to be manipulated using genetic modification. This review aims to highlight the recent studies that have been carried out using marine lipases for the enrichment of omega-3, to provide insight into future directions. Overall, the covalent bond-based lipase immobilization to various support materials appears most promising; however, greener and less expensive options need to be strengthened.


Sujet(s)
Organismes aquatiques , Compléments alimentaires , Acides gras omega-3 , Triacylglycerol lipase , Triacylglycerol lipase/métabolisme , Triacylglycerol lipase/composition chimique , Acides gras omega-3/composition chimique , Humains , Animaux , Huiles de poisson/composition chimique
3.
Int J Mol Sci ; 25(13)2024 Jul 02.
Article de Anglais | MEDLINE | ID: mdl-39000384

RÉSUMÉ

The incidence of nonalcoholic fatty liver disease (NAFLD), or metabolic dysfunction-associated fatty liver disease (MAFLD), is increasing in adults and children. Unfortunately, effective pharmacological treatments remain unavailable. Single nucleotide polymorphisms (SNPs) in the patatin-like phospholipase domain-containing protein (PNPLA3 I148M) have the most significant genetic association with the disease at all stages of its progression. A roadblock to identifying potential treatments for PNPLA3-induced NAFLD is the lack of a human cell platform that recapitulates the PNPLA3 I148M-mediated onset of lipid accumulation. Hepatocyte-like cells were generated from PNPLA3-/- and PNPLA3I148M/M-induced pluripotent stem cells (iPSCs). Lipid levels were measured by staining with BODIPY 493/503 and were found to increase in PNPLA3 variant iPSC-derived hepatocytes. A small-molecule screen identified multiple compounds that target Src/PI3K/Akt signaling and could eradicate lipid accumulation in these cells. We found that drugs currently in clinical trials for cancer treatment that target the same pathways also reduced lipid accumulation in PNPLA3 variant cells.


Sujet(s)
Hépatocytes , Cellules souches pluripotentes induites , Triacylglycerol lipase , Protéines membranaires , Stéatose hépatique non alcoolique , Hépatocytes/métabolisme , Humains , Cellules souches pluripotentes induites/métabolisme , Protéines membranaires/métabolisme , Protéines membranaires/génétique , Stéatose hépatique non alcoolique/métabolisme , Stéatose hépatique non alcoolique/génétique , Stéatose hépatique non alcoolique/anatomopathologie , Stéatose hépatique non alcoolique/étiologie , Triacylglycerol lipase/métabolisme , Triacylglycerol lipase/génétique , Transduction du signal , Métabolisme lipidique , Polymorphisme de nucléotide simple , Acyltransferases , Calcium-independent phospholipase A2
4.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 41(7): 840-843, 2024 Jul 10.
Article de Chinois | MEDLINE | ID: mdl-38946369

RÉSUMÉ

OBJECTIVE: To explore the clinical phenotype and genetic basis of a child with Neutral lipid storage disease with myopathy (NLSDM). METHODS: A child who was admitted to the First Affiliated Hospital of Zhengzhou University in February 2021 for a history of elevated creatine kinase (CK) for over 2 months was selected as the study subject. Clinical and laboratory examinations were carried out, and the child was subjected to whole exome sequencing. Candidate variants were validated by Sanger sequencing of her family members. RESULTS: The patient, a 9-year-old female, had exhibited weakness in the lower limbs, elevated CK level, and refractory cardiomyotrophy. Genetic testing revealed that she has harbored c.32C>G (p.S11W) and c.516C>G (p.N172K) compound heterozygous variants of the PNPLA2 gene, which were respectively inherited from her mother and father. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), both variants were rated as likely pathogenic (PM1+PM2_Supporting+PP3+PP4). CONCLUSION: The c.32C>G (p.S11W) and c.516C>G (p.N172K) compound heterozygous variants of the PNPLA2 gene probably underlay the myasthenia gravis and elevated creatine kinase in this child.


Sujet(s)
Triacylglycerol lipase , Erreurs innées du métabolisme lipidique , Maladies musculaires , Humains , Femelle , Enfant , Maladies musculaires/génétique , Erreurs innées du métabolisme lipidique/génétique , Triacylglycerol lipase/génétique , Mutation , Dépistage génétique , , Creatine kinase/sang , Pedigree , Phénotype , Acyltransferases
5.
Nat Commun ; 15(1): 5773, 2024 Jul 10.
Article de Anglais | MEDLINE | ID: mdl-38982057

RÉSUMÉ

Harnessing the power of cell biocatalysis for sustainable chemical synthesis requires rational integration of living cells with the modern synthetic catalysts. Here, we develop silica-tiling strategy that constructs a hierarchical, inorganic, protocellular confined nanospace around the individual living cell to accommodate molecularly accessible abiotic catalytic sites. This empowers the living microorganisms for new-to-nature chemical synthesis without compromising the cellular regenerative process. Yeast cell, a widely used biocatalyst, is upgraded via highly controlled self-assembly of 2D-bilayer silica-based catalytic modules on cell surfaces, opening the avenues for diverse chemobiotic reactions. For example, combining [AuPt]-catalyzed NADH regeneration, light-induced [Pd]-catalyzed C-C cross-coupling or lipase-catalyzed esterification reactions-with the natural ketoreductase activity inside yeast cell. The conformal silica bilayer provides protection while allowing proximity to catalytic sites and preserving natural cell viability and proliferation. These living nanobiohybrids offer to bridge cell's natural biocatalytic capabilities with customizable heterogeneous metal catalysis, enabling programmable reaction sequences for sustainable chemical synthesis.


Sujet(s)
Biocatalyse , Saccharomyces cerevisiae , Silice , Silice/composition chimique , Saccharomyces cerevisiae/métabolisme , Catalyse , NAD/métabolisme , Triacylglycerol lipase/métabolisme , Palladium/composition chimique , Estérification
6.
Appl Microbiol Biotechnol ; 108(1): 413, 2024 Jul 10.
Article de Anglais | MEDLINE | ID: mdl-38985324

RÉSUMÉ

Environmental concerns arising from the increasing use of polluting plastics highlight polylactic acid (PLA) as a promising eco-friendly alternative. PLA is a biodegradable polyester that can be produced through the fermentation of renewable resources. Together with its excellent properties, suitable for a wide range of applications, the use of PLA has increased significantly over the years and is expected to further grow. However, insufficient degradability under natural conditions emphasizes the need for the exploration of biodegradation mechanisms, intending to develop more efficient techniques for waste disposal and recycling or upcycling. Biodegradation occurs through the secretion of depolymerizing enzymes, mainly proteases, lipases, cutinases, and esterases, by various microorganisms. This review focuses on the enzymatic degradation of PLA and presents different enzymes that were isolated and purified from natural PLA-degrading microorganisms, or recombinantly expressed. The review depicts the main characteristics of the enzymes, including recent advances and analytical methods used to evaluate enantiopurity and depolymerizing activity. While complete degradation of solid PLA particles is still difficult to achieve, future research and improvement of enzyme properties may provide an avenue for the development of advanced procedures for PLA degradation and upcycling, utilizing its building blocks for further applications as envisaged by circular economy principles. KEY POINTS: • Enzymes can be promisingly utilized for PLA upcycling. • Natural and recombinant PLA depolymerases and methods for activity evaluation are summarized. • Approaches to improve enzymatic degradation of PLA are discussed.


Sujet(s)
Dépollution biologique de l'environnement , Polyesters , Polyesters/métabolisme , Carboxylic ester hydrolases/métabolisme , Carboxylic ester hydrolases/génétique , Triacylglycerol lipase/métabolisme , Esterases/métabolisme , Bactéries/enzymologie , Bactéries/métabolisme , Peptide hydrolases/métabolisme
7.
Physiol Rep ; 12(13): e16097, 2024 Jul.
Article de Anglais | MEDLINE | ID: mdl-38955666

RÉSUMÉ

Latent associations between low serum amylase and reduced plasma insulin levels and increased adiposity have been described previously in a small study of asymptomatic middle-aged humans. In the present study, we sought to determine the nature of such changes during the longitudinal progression from metabolically normal to overt type 2 diabetes mellitus (T2DM) in nonhuman primates (NHPs), a disease that appears to be the same in both pathophysiology and underlying mechanisms as that which most commonly develops in middle-aged adult humans. Amylase and lipase levels were characterized in 157 unrelated adult rhesus monkeys (Macaca mulatta); 38% developed T2DM while under study. In all monkeys, multivariable linear regression analysis revealed that amylase could be negatively predicted by % body fat (ß -0.29; p = 0.002), age (ß -0.27; p = 0.005), and HbA1c (ß -0.18; p = 0.037). Amylase levels were positively predicted by lipase levels (ß = 0.19; p = -0.024) in all NHPs included in the study. Amylase was significantly lower in NHPs with metabolic syndrome (p < 0.001), prediabetes (PreDM) (p < 0.001), and T2DM (p < 0.001) compared to metabolically normal adult NHPs. Lipase increased in NHPs with PreDM (p = 0.005) and T2DM (p = 0.04) compared to normal NHPs. This is the first longitudinal study of any species, including humans, to show the dynamics of amylase and lipase during the metabolic progression from normal to metabolic syndrome, to PreDM and then to overt T2DM. The extraordinary similarity between humans and monkeys in T2DM, in pancreatic pathophysiology and in metabolic functions give these findings high translational value.


Sujet(s)
Amylases , Diabète de type 2 , Triacylglycerol lipase , Macaca mulatta , Syndrome métabolique X , Animaux , Diabète de type 2/sang , Diabète de type 2/métabolisme , Triacylglycerol lipase/sang , Triacylglycerol lipase/métabolisme , Mâle , Syndrome métabolique X/sang , Syndrome métabolique X/métabolisme , Études longitudinales , Amylases/sang , Amylases/métabolisme , Femelle
8.
Med Microbiol Immunol ; 213(1): 16, 2024 Jul 20.
Article de Anglais | MEDLINE | ID: mdl-39033094

RÉSUMÉ

Melioidosis is a severe infectious disease caused by Burkholderia pseudomallei, an intracellular pathogen with a high mortality rate and significant antibiotic resistance. The high mortality rate and resistance to antibiotics have drawn considerable attention from researchers studying melioidosis. This study evaluated the effects of various concentrations (75, 50, and 25 µg/mL) of promethazine hydrochloride (PTZ), a potent antihistamine, on biofilm formation and lipase activity after 24 h of exposure to B. thailandensis E264. A concentration-dependent decrease in both biofilm biomass and lipase activity was observed. RT-PCR analysis revealed that PTZ treatment not only made the biofilm structure loose but also reduced the expression of btaR1, btaR2, btaR3, and scmR. Single gene knockouts of quorum sensing (QS) receptor proteins (∆btaR1, ∆btaR2, and ∆btaR3) were successfully constructed. Deletion of btaR1 affected biofilm formation in B. thailandensis, while deletion of btaR2 and btaR3 led to reduced lipase activity. Molecular docking and biological performance results demonstrated that PTZ inhibits biofilm formation and lipase activity by suppressing the expression of QS-regulated genes. This study found that repositioning PTZ reduced biofilm formation in B. thailandensis E264, suggesting a potential new approach for combating melioidosis.


Sujet(s)
Biofilms , Burkholderia , Repositionnement des médicaments , Prométhazine , Biofilms/effets des médicaments et des substances chimiques , Biofilms/croissance et développement , Burkholderia/effets des médicaments et des substances chimiques , Burkholderia/physiologie , Burkholderia/génétique , Prométhazine/pharmacologie , Simulation de docking moléculaire , Antibactériens/pharmacologie , Triacylglycerol lipase/métabolisme , Triacylglycerol lipase/génétique , Régulation de l'expression des gènes bactériens/effets des médicaments et des substances chimiques , Protéines bactériennes/génétique , Protéines bactériennes/métabolisme , Humains , Détection du quorum/effets des médicaments et des substances chimiques
9.
Langmuir ; 40(29): 15293-15300, 2024 Jul 23.
Article de Anglais | MEDLINE | ID: mdl-39007240

RÉSUMÉ

Controlling physicochemical processes that drive changes in supramolecular aggregates is an important objective toward creating artificial soft micro- and nanomachines. Previous research explored the morphology control of membrane-based materials subjected to externally imposed chemical stimuli. Here, we modulate the microscale morphology of pH-responsive assemblies by using biocatalysis to internally generate changes in global pH. Catalytic reactions offer flexibility in the mechanism and rate at which stimuli are introduced to responsive assemblies, ultimately enabling precision and control over size and morphology. We observed, by dynamic light scattering and fluorescence microscopy, substantial microscale differences between assemblies subjected to manually titrated pH changes compared to biocatalytically activated pH changes, including the growth of giant vesicles from micelles. Coarse-grained molecular dynamics simulations of these metastable self-assembled structures provided insight into the thermodynamics and kinetics of the preferred structures. These results demonstrate the feasibility of using biocatalytic reactions to modulate the size and morphology of supramolecular assemblies, from micelles to giant vesicles.


Sujet(s)
Biocatalyse , Micelles , Concentration en ions d'hydrogène , Simulation de dynamique moléculaire , Triacylglycerol lipase/composition chimique , Triacylglycerol lipase/métabolisme , Cinétique , Thermodynamique
10.
Methods Mol Biol ; 2816: 253-263, 2024.
Article de Anglais | MEDLINE | ID: mdl-38977604

RÉSUMÉ

Lipids are compounds involved in many biologic functions including cell structure, metabolism, energy storage and are involved in signaling. A prominent lipid in these functions is cholesterol. Cholesterol also plays a part in the signaling of melanocytes, which contain melanosomes. The maturation of these melanosomes happens during melanocyte growth. The deficit of melanogenesis or melanosome maturation is associated with ocular albinism in the eye. Aberrations of melanosome maturation are also associated with pigment dispersion syndrome. Albinism and pigment dispersion manifestations are systemic. Both melanogenesis and melanocyte maturation are affected by cholesterol metabolism. Cholesterol signaling is a part of many pathways in the body, and evaluating these signals can have implications in systemic disease processes of melanogenesis and melanosome maturation, like ocular albinism and pigment dispersion. Cholesterol is carried by lipoprotein particles. Low-density lipoprotein (LDL) is usually the transport vehicle for cholesterol to reach tissues and organelles. The LDL uptake on cells often sends out a cascade of internal signaling within the cells. We describe here LDL signaling related to lipase activity changes using enzymatic methods with a kit. We describe analyses of cholesterol esters and free cholesterol with liquid chromatography and gas chromatography with or in tandem with mass spectrometry (GC-MS and LC-MS/MS). These analyses will provide insight into melanosome maturation and melanogenesis. The methods described here are applicable to all melanocytes within the body of a model mammalian organism.


Sujet(s)
Cholestérol , Iris , Mélanocytes , Transduction du signal , Mélanocytes/métabolisme , Humains , Cholestérol/métabolisme , Iris/métabolisme , Lipoprotéines/métabolisme , Mélanosomes/métabolisme , Lipoprotéines LDL/métabolisme , Spectrométrie de masse en tandem/méthodes , Chromatographie gazeuse-spectrométrie de masse/méthodes , Chromatographie en phase liquide/méthodes , Triacylglycerol lipase/métabolisme , Mélanines/métabolisme , Cholestérol ester/métabolisme
11.
Nutrients ; 16(14)2024 Jul 14.
Article de Anglais | MEDLINE | ID: mdl-39064713

RÉSUMÉ

(1) Background: Proglucagon-derived peptides (PDGPs) including glucagon (Gcg), GLP-1, and GLP-2 regulate lipid metabolism in the liver, adipocytes, and intestine. However, the mechanism by which PGDPs participate in alterations in lipid metabolism induced by high-fat diet (HFD) feeding has not been elucidated. (2) Methods: Mice deficient in PGDP (GCGKO) and control mice were fed HFD for 7 days and analyzed, and differences in lipid metabolism in the liver, adipose tissue, and duodenum were investigated. (3) Results: GCGKO mice under HFD showed lower expression levels of the genes involved in free fatty acid (FFA) oxidation such as Hsl, Atgl, Cpt1a, Acox1 (p < 0.05), and Pparα (p = 0.05) mRNA in the liver than in control mice, and both FFA and triglycerides content in liver and adipose tissue weight were lower in the GCGKO mice. On the other hand, phosphorylation of hormone-sensitive lipase (HSL) in white adipose tissue did not differ between the two groups. GCGKO mice under HFD exhibited lower expression levels of Pparα and Cd36 mRNA in the duodenum as well as increased fecal cholesterol contents compared to HFD-controls. (4) Conclusions: GCGKO mice fed HFD exhibit a lesser increase in hepatic FFA and triglyceride contents and adipose tissue weight, despite reduced ß-oxidation in the liver, than in control mice. Thus, the absence of PGDP prevents dietary-induced fatty liver development due to decreased lipid uptake in the intestinal tract.


Sujet(s)
Antigènes CD36 , Alimentation riche en graisse , Absorption intestinale , Métabolisme lipidique , Foie , Souris knockout , Récepteur PPAR alpha , Proglucagon , Animaux , Mâle , Alimentation riche en graisse/effets indésirables , Récepteur PPAR alpha/métabolisme , Récepteur PPAR alpha/génétique , Foie/métabolisme , Proglucagon/métabolisme , Proglucagon/génétique , Antigènes CD36/métabolisme , Antigènes CD36/génétique , Souris , Sterol Esterase/métabolisme , Sterol Esterase/génétique , Triglycéride/métabolisme , Souris de lignée C57BL , Acide gras libre/métabolisme , Glucagon-like peptide 1/métabolisme , Duodénum/métabolisme , Carnitine O-palmitoyltransferase/métabolisme , Carnitine O-palmitoyltransferase/génétique , Tissu adipeux/métabolisme , Matières grasses alimentaires , Glucagon-like peptide 2/métabolisme , Acyltransferases , Triacylglycerol lipase
12.
Molecules ; 29(14)2024 Jul 10.
Article de Anglais | MEDLINE | ID: mdl-39064846

RÉSUMÉ

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.


Sujet(s)
Triacylglycerol lipase , Triacylglycerol lipase/métabolisme , Triacylglycerol lipase/composition chimique , Stéréoisomérie , Protéines fongiques/métabolisme , Biocatalyse , Pipérazines/composition chimique , Pipérazines/synthèse chimique , Enzymes immobilisées/composition chimique , Solvants/composition chimique , Structure moléculaire
13.
Molecules ; 29(14)2024 Jul 12.
Article de Anglais | MEDLINE | ID: mdl-39064875

RÉSUMÉ

This study presents the potential role of deep eutectic solvents (DESs) in a lipase-catalyzed hydrolysis reaction as a co-solvent in an aqueous solution given by a phosphate buffer. Ammonium salts, such as choline chloride, were paired with hydrogen bond donors, such as urea, 1,2,3-propanetriol, and 1,2 propanediol. The hydrolysis of p-nitrophenyl laureate was carried out with the lipase Candida antarctica Lipase B (CALB) as a reaction model to evaluate the solvent effect and tested in different DES/buffer phosphate mixtures at different % w/w. The results showed that two mixtures of different DES at 25 % w/w were the most promising solvents, as this percentage enhanced the activities of CALB, as evidenced by its higher catalytic efficiency (kcatKM). The solvent analysis shows that the enzymatic reaction requires a reaction media rich in water molecules to enable hydrogen-bond formation from the reaction media toward the enzymatic reaction, suggesting a better interaction between the substrate and the enzyme-active site. This interaction could be attributed to high degrees of freedom influencing the enzyme conformation given by the reaction media, suggesting that CALB acquires a more restrictive structure in the presence of DES or the stabilized network given by the hydrogen bond from water molecules in the mixture improves the enzymatic activity, conferring conformational stability by solvent effects. This study offers a promising approach for applications and further perspectives on genuinely green industrial solvents.


Sujet(s)
Solvants eutectiques profonds , Protéines fongiques , Liaison hydrogène , Triacylglycerol lipase , Eau , Triacylglycerol lipase/composition chimique , Triacylglycerol lipase/métabolisme , Eau/composition chimique , Solvants eutectiques profonds/composition chimique , Protéines fongiques/composition chimique , Catalyse , Hydrolyse , Solvants/composition chimique , Biocatalyse , Cinétique
14.
Int J Mol Sci ; 25(14)2024 Jul 19.
Article de Anglais | MEDLINE | ID: mdl-39063171

RÉSUMÉ

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.


Sujet(s)
Stabilité enzymatique , Geobacillus , Triacylglycerol lipase , Protéines recombinantes , Solvants , Geobacillus/enzymologie , Geobacillus/génétique , Triacylglycerol lipase/génétique , Triacylglycerol lipase/composition chimique , Triacylglycerol lipase/métabolisme , Triacylglycerol lipase/isolement et purification , Solvants/composition chimique , Régions antarctiques , Protéines recombinantes/composition chimique , Protéines recombinantes/métabolisme , Protéines recombinantes/génétique , Concentration en ions d'hydrogène , Protéines bactériennes/génétique , Protéines bactériennes/composition chimique , Protéines bactériennes/métabolisme , Cinétique , Spécificité du substrat , Température , Escherichia coli/génétique , Escherichia coli/métabolisme
15.
Mol Biol Rep ; 51(1): 845, 2024 Jul 23.
Article de Anglais | MEDLINE | ID: mdl-39042259

RÉSUMÉ

BACKGROUND: This study examines genetic variations in CYP2E1 (rs6413432, rs3813867), GCKR (rs780094, rs1260326), and PNPLA3 (rs738409) among Turkish patients to assess their influence on nonalcoholic steatohepatitis. METHODS: Allele and genotype frequencies were compared between 245 NASH patients and 120 healthy controls using SNP genotyping via polymerase chain reaction-restriction fragment length polymorphism. Additionally, the deviation of the observed genotype frequencies from Hardy-Weinberg proportion was examined. RESULTS: No significant differences were found in the allelic and genotypic distributions of rs6413432, rs3813867, and rs780094 between NASH patients and healthy controls. However, significant disparities were noted for rs1260326 and rs738409. Gender and age-specific distributions showed no notable differences. The only observed deviation from Hardy-Weinberg proportion was in the genotype frequency of rs738409. CONCLUSIONS: Variants in GCKR (rs1260326) and PNPLA3 (rs738409) are significantly associated with increased NASH risk in the Turkish population, with the rs738409 variant potentially playing a more prominent role in NASH development.


Sujet(s)
Protéines adaptatrices de la transduction du signal , Cytochrome P-450 CYP2E1 , Fréquence d'allèle , Prédisposition génétique à une maladie , Génotype , Triacylglycerol lipase , Protéines membranaires , Stéatose hépatique non alcoolique , Polymorphisme de nucléotide simple , Humains , Mâle , Femelle , Turquie , Triacylglycerol lipase/génétique , Polymorphisme de nucléotide simple/génétique , Stéatose hépatique non alcoolique/génétique , Adulte d'âge moyen , Adulte , Protéines membranaires/génétique , Fréquence d'allèle/génétique , Cytochrome P-450 CYP2E1/génétique , Protéines adaptatrices de la transduction du signal/génétique , Allèles , Études cas-témoins , Sujet âgé , Acyltransferases , Calcium-independent phospholipase A2
16.
Appl Microbiol Biotechnol ; 108(1): 399, 2024 Jun 29.
Article de Anglais | MEDLINE | ID: mdl-38951177

RÉSUMÉ

Dehydroepiandrosterone (DHEA) has a promising market due to its capacity to regulate human hormone levels as well as preventing and treating various diseases. We have established a chemical esterification coupled biocatalytic-based scheme by lipase-catalyzed 4-androstene-3,17-dione (4-AD) hydrolysis to obtain the intermediate product 5-androstene-3,17-dione (5-AD), which was then asymmetrically reduced by a ketoreductase from Sphingomonas wittichii (SwiKR). Co-enzyme required for KR is regenerated by a glucose dehydrogenase (GDH) from Bacillus subtilis. This scheme is more environmentally friendly and more efficient than the current DHEA synthesis pathway. However, a significant amount of 4-AD as by-product was detected during the catalytic process. Focused on the control of by-products, we investigated the source of 4-AD and identified that it is mainly derived from the isomerization activity of SwiKR and GDH. Increasing the proportion of glucose in the catalytic system as well as optimizing the catalytic conditions drastically reduced 4-AD from 24.7 to 6.5% of total substrate amount, and the final yield of DHEA achieved 40.1 g/L. Furthermore, this is the first time that both SwiKR and GDH have been proved to be promiscuous enzymes with dehydrogenase and ketosteroid isomerase (KSI) activities, expanding knowledge of the substrate diversity of the short-chain dehydrogenase family enzymes. KEY POINTS: • A strategy of coupling lipase, ketoreductase, and glucose dehydrogenase in producing DHEA from 4-AD • Both SwiKR and GDH are identified with ketosteroid isomerase activity. • Development of catalytic strategy to control by-product and achieve highly selective DHEA production.


Sujet(s)
Déhydroépiandrostérone , Triacylglycerol lipase , Sphingomonas , Déhydroépiandrostérone/métabolisme , Triacylglycerol lipase/métabolisme , Sphingomonas/enzymologie , Sphingomonas/métabolisme , Biocatalyse , Bacillus subtilis/enzymologie , Bacillus subtilis/métabolisme , Bacillus subtilis/génétique , Glucose 1-dehydrogenase/métabolisme , Glucose 1-dehydrogenase/génétique , Androstènedione/métabolisme , Androstènedione/biosynthèse , Hydrolyse
17.
PLoS One ; 19(6): e0298253, 2024.
Article de Anglais | MEDLINE | ID: mdl-38843179

RÉSUMÉ

Stunting is caused by various factors, including low nutritional intake in the first two years of life. This study aimed to investigate the differences in sociodemographic factors and mineral, vitamin, and enzyme parameters in mothers associated with the occurrence of stunting in children. We conducted a cross-sectional study from September to November 2020 on North Sumatra Island, Indonesia. The data collected included sociodemographic characteristics, pregnancy history, birth history, food intake, and laboratory examinations, including measurements of calcium, iron, zinc, vitamin D, pancreatic amylase, and serum lipase levels. This study included 50 healthy mothers aged 18-50 years old with children aged 2 to 60 months. There was a significant difference in serum calcium levels between the groups of mothers of children with normal and stunted growth (p = 0.03, mean difference±standard error (SE) = 0.23±0.12, 95% CI: 0.19-0.45). All of the study subjects were categorized as vitamin D deficient. The mean lipase level in the group of mothers of children with stunted growth was significantly lower than that in the group of mothers of children with normal growth (p = 0.02, mean difference±SE = 4.34±1.83, 95% CI: 0.62-8.06). The conclusion was that serum lipase levels were significantly lower in mothers of children with stunted growth compared to mothers of children with normal growth. Serum lipase levels this low are likely to indicate that a mother is unable to meet her child's calcium needs during pregnancy, increasing the child's risk of stunted growth.


Sujet(s)
Calcium , Troubles de la croissance , Triacylglycerol lipase , Humains , Femelle , Indonésie/épidémiologie , Grossesse , Études transversales , Adulte , Calcium/sang , Triacylglycerol lipase/sang , Troubles de la croissance/sang , Troubles de la croissance/épidémiologie , Adolescent , Nourrisson , Enfant d'âge préscolaire , Jeune adulte , Mères , Adulte d'âge moyen , Mâle , Vitamine D/sang , Vitamine D/analogues et dérivés , Carence en vitamine D/sang , Carence en vitamine D/épidémiologie
18.
Carbohydr Polym ; 341: 122353, 2024 Oct 01.
Article de Anglais | MEDLINE | ID: mdl-38876720

RÉSUMÉ

The use of Pickering emulsions for biocatalysis is gaining increased attention. However, the extensive application is greatly limited due to the enzyme inactivation. Herein, a biocatalytic Pickering emulsion with high-performance utilizing cellulose nanocrystals immobilized lipases (CNCs-Lps) particles as stabilizer is advanced and applied for the synthesis of Vitamin E nicotinate. CNCs-Lps display high activity and reusability due to the construction of biocatalytic microreactor in the O/W emulsion system. The yield of vitamin E nicotinate ester reached up to 83 %. More importantly, the CNCs-Lps can be reused due to the similar principles to microreactors in Pickering emulsions. Reusability test showed that the CNCs-Lps could be recovered from the emulsion system by centrifugation and the yield of vitamin E nicotinate retains 78 % of initial value after five cycles, demonstrating overwhelming advantage than the fair counterpart with free lipases.


Sujet(s)
Biocatalyse , Cellulose , Émulsions , Enzymes immobilisées , Triacylglycerol lipase , Nanoparticules , Cellulose/composition chimique , Émulsions/composition chimique , Triacylglycerol lipase/composition chimique , Triacylglycerol lipase/métabolisme , Nanoparticules/composition chimique , Enzymes immobilisées/composition chimique , Enzymes immobilisées/métabolisme , Vitamine E/composition chimique
19.
Nat Commun ; 15(1): 4847, 2024 Jun 06.
Article de Anglais | MEDLINE | ID: mdl-38844467

RÉSUMÉ

The I148M variant of PNPLA3 is closely associated with hepatic steatosis. Recent evidence indicates that the I148M mutant functions as an inhibitor of PNPLA2/ATGL-mediated lipolysis, leaving the role of wild-type PNPLA3 undefined. Despite showing a triglyceride hydrolase activity in vitro, PNPLA3 has yet to be established as a lipase in vivo. Here, we show that PNPLA3 preferentially hydrolyzes polyunsaturated triglycerides, mobilizing polyunsaturated fatty acids for phospholipid desaturation and enhancing hepatic secretion of triglyceride-rich lipoproteins. Under lipogenic conditions, mice with liver-specific knockout or acute knockdown of PNPLA3 exhibit aggravated liver steatosis and reduced plasma VLDL-triglyceride levels. Similarly, I148M-knockin mice show decreased hepatic triglyceride secretion during lipogenic stimulation. Our results highlight a specific context whereby the wild-type PNPLA3 facilitates the balance between hepatic triglyceride storage and secretion, and suggest the potential contribution of a loss-of-function by the I148M variant to the development of fatty liver disease in humans.


Sujet(s)
Acides gras insaturés , Triacylglycerol lipase , Lipoprotéines VLDL , Foie , Souris knockout , Triglycéride , Animaux , Triacylglycerol lipase/métabolisme , Triacylglycerol lipase/génétique , Foie/métabolisme , Triglycéride/métabolisme , Souris , Lipoprotéines VLDL/métabolisme , Humains , Acides gras insaturés/métabolisme , Mâle , Stéatose hépatique/métabolisme , Stéatose hépatique/génétique , Souris de lignée C57BL , Lipolyse , Protéines membranaires/métabolisme , Protéines membranaires/génétique , Acyltransferases , Calcium-independent phospholipase A2
20.
BMC Microbiol ; 24(1): 209, 2024 Jun 14.
Article de Anglais | MEDLINE | ID: mdl-38877423

RÉSUMÉ

Fungi can spoil the majority of baked products. Spoilage of cake during storage is commonly associated with fungi. Therefore, this study aimed to assess the quality of different types of cakes sold in the market. The most predominant fungal genera in the tested cake samples (14 samples) were Aspergillus spp., and Penicillium spp. On Potato Dextrose Agar (PDA), the medium fungal total count was 43.3 colonies /g. Aspergillus was the most dominant genus and was isolated from six samples of cake. Aspergillus was represented by 3 species namely, A. flavus, A. niger, and A. nidulans, represented by 13.32, 19.99, and 3.33 colonies /g respectively. On Malt Extract Agar (MEA) Medium, the fungal total count was 123.24 colonies / g. Aspergillus was the most dominant isolated genus from 11 samples of cake and was represented by 5 species, namely, A. flavus, A. niger, A. ochraceous, A. terreus, and A. versicolor (26. 65, 63.29, 3.33, 6.66, and 3.33 colonies / g , respectively). Twenty-four isolates (88.88 %) of the total tested twenty-seven filamentous fungi showed positive results for amylase production. Ten isolates (37.03%) of the total tested filamentous fungi showed positive results for lipase production, and finally eleven isolates (40.74 %) of the total fungal isolates showed positive results for protease production. Aflatoxins B1, B2, G1, G2, and ochratoxin A were not detected in fourteen collected samples of cake. In this study, clove oil was the best choice overpeppermint oil and olive oil for preventing mold development when natural agents were compared. It might be due to the presence of a varietyof bioactive chemical compounds in clove oil, whose major bioactive component is eugenol, which acts as an antifungal reagent. Therefore, freshly baked cake should be consumed within afew days to avoid individuals experiencing foodborne illnesses.


Sujet(s)
Microbiologie alimentaire , Champignons , Mycotoxines , Champignons/isolement et purification , Champignons/classification , Champignons/enzymologie , Champignons/génétique , Mycotoxines/analyse , Aspergillus/isolement et purification , Aspergillus/enzymologie , Penicillium/isolement et purification , Penicillium/enzymologie , Contamination des aliments/analyse , Aflatoxines/analyse , Triacylglycerol lipase/métabolisme , Amylases/métabolisme , Amylases/analyse
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