Improved Enzymatic Properties of Chitosanase CsnMY002 from Bacillus subtilis via Computational Design

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(4), P. 1588 - 1588

Published: Feb. 13, 2025

Chitooligosaccharides (COSs) are a class of functional carbohydrates with significant application prospects in food and medicine. Chitosanase CsnMY002 from the GH46 family has been used to prepare COS controlled degrees polymerization. To enhance industrial applicability CsnMY002, molecular dynamics (MD) simulations were applied investigate structure-property relationship. Guided by simulation results, beneficial mutants screened through synergistic strategy using residue-folding free energy calculation consensus sequence analysis. Iterative combinations constructed mutant Mut6 (A49G/K70A/S84A/N89G/D199R/N221G) significantly improved thermal stability, which had half-life (t1/2 value) at 55 °C 75 that was 1.80 1.62 times higher than wild type, respectively. A highly active mutant, Mut2, created, exhibiting 1.52 catalytic efficiency type. An MD analysis suggested enzymatic properties correlated changes dynamic behaviours enzyme structure. This study generated more suitable variants for production provided comprehensive optimization other enzymes potential.

Language: Английский

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Thermal stability enhancement of erythromycin esterase C from Enterobacter hormaechei through a combinatorial greedy accumulated strategy

International Journal of Biological Macromolecules, Journal Year: 2025, Volume and Issue: 305, P. 141207 - 141207

Published: Feb. 17, 2025

Language: Английский

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High-efficiency secretion expression of cellobiose 2-epimerase in Escherichia coli and its applications

International Journal of Biological Macromolecules, Journal Year: 2025, Volume and Issue: unknown, P. 142205 - 142205

Published: March 1, 2025

Language: Английский

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Computer-Aided Molecular Modifications for Enhanced Activity and Thermal Stability of d-Allulose 3-Epimerase

Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 26, 2025

D-Allulose 3-epimerase (DAE) can convert D-fructose into D-allulose, but its lower activity and poor thermal stability restrict application in industrial production. This study utilized a semi-rational design specific site mutations to enhance the enzyme of DAE from Clostridium scindens ATCC 35704 (Csc-DAE) at same time. Notably, activities six mutants (V22I, A25V, Q46I, C212N, G165A, G267D) orderly superimposed double-point were increased by 1.08–1.31 1.28–1.64 times that wild type (553.7 U/mg), respectively. In addition, A25V/C212N showed good stability, optimum temperature (Topt) was 65 °C half-life (t1/2) 3.997 h 60 °C, which 5 5.76 times, The conversion ratio reached 30.62 31.22% 70 respectively, structural changes analyzed using molecular docking. These studies provide beneficial mutation for applications.

Language: Английский

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Biocatalytic Preparation of High-Purity DHA-Enriched Partial Glycerides through Semirational Design and Modification of Lipase Lip1897

Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 31, 2025

High-purity docosahexaenoic acid (DHA) partial glycerides (PG) supplements have promising market prospects. In this study, a novel lipolytic enzyme family I lipase Lip1897, derived from Streptomyces thermodiastaticus, was identified and modified to be applied the enzymatic synthesis of high-purity (DHA-PG). Lip1897 displayed optimal activity good stability at 55 °C pH 5.0. Further study found that could catalyze hydrolysis DHA triglycerides (TG) ethyl esters (EE), esterification DHA, glycerolysis (DHA-EE), which had prospects for industrial application. To enhance catalytic selectivity semirational design modification based on structural analysis were carried out, mutant Lip1897-H106W showed 1.86-fold increase in activity. The molecular docking results indicated bound substrate DHA-EE lower energy with more stable conformation. Under solvent-free system, employed DHA-PG, achieving conversion rate 95.02% high DHA-PG yield 70.85%. efficient preparing realized. This research provides reference enhancing efficiency specific configurational functional lipid biosynthesis through discovery modification.

Language: Английский

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Immobilization of fungal laccase on peanut shell carriers

Biocatalysis and Agricultural Biotechnology, Journal Year: 2025, Volume and Issue: unknown, P. 103581 - 103581

Published: April 1, 2025

Language: Английский

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