Computer-assisted enzyme cocktails enhance fermentation by overcoming toxic inhibitors from pretreatment processes

Bioresource Technology, Год журнала: 2025, Номер 419, С. 132076 - 132076

Опубликована: Янв. 18, 2025

Язык: Английский

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Biochemical and Structural Characterization of a Highly Glucose-Tolerant β-Glucosidase from the Termite Reticulitermes perilucifugus

International Journal of Molecular Sciences, Год журнала: 2025, Номер 26(7), С. 3118 - 3118

Опубликована: Март 28, 2025

The enzymatic hydrolysis of lignocellulose is often hindered by the glucose-mediated inhibition β-glucosidases, a major bottleneck in industrial cellulose degradation. Identifying novel glucose-tolerant β-glucosidases essential for enhancing saccharification efficiency. In this study, we cloned and heterologously expressed β-glucosidase, RpBgl8, from termite Reticulitermes perilucifugus Escherichia coli. Sequence structural analyses classified RpBgl8 as glycoside hydrolase family 1 enzyme. purified enzyme exhibited optimal activity at 45 °C pH 7.0, with broad stability across 4.0-8.0. Notably, demonstrated high tolerance to lignocellulose-derived inhibitors organic solvents, maintaining 100% 15% ethanol. Furthermore, outstanding glucose tolerance, retaining 2.5 M 82% 4.0 glucose-outperforming most previously reported β-glucosidases. A analysis revealed narrow, hydrophobic substrate pocket, residue F124 glycone-binding site critical minimizing accumulation. F124W mutation significantly reduced confirming that interactions active mitigate inhibition. These findings establish promising candidate high-solid biomass processing simultaneous fermentation applications, highlighting termites underexplored sources biocatalysts unique potential.

Язык: Английский

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Enhancing the thermostability of phospholipase C by structural-based proline incorporation to improve its degumming performance

Опубликована: Апрель 18, 2025

Thermostability can be improved by introducing prolines into targeted sites, enhancing enzyme performance in specific reactions. In our present study, a novel fungal phospholipase C derived from Talaromyces islandicus (TiPLC) was first heterologously expressed Pichia pastoris and biochemically characterized. Given the poor thermal stability of TiPLC, structure-based proline incorporation strategy used to enhance its thermostability further. Two single-site (E92P A375P) mutants were selected seven designs, exhibiting while retaining wild-type's basic properties (optimum reaction pH temperature). Compared wild-type, t_1/2 E92P A375P under 40 ℃ extended 1.62 1.27 times, respectively. Meanwhile, exhibited 20% increase activity using p-NPPC as substrate. Moreover, double mutant E92P-A375P 2.43 times enhancement compared wild-type. Results oil degumming experiment further confirmed that significantly with reduction residual phosphorus 78 ppm, for 131 ppm same reaction. Molecular simulations indicated 92 375 sites rigidity partial flexible regions, thus contributing increased thermostability.

Язык: Английский

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Unlocking the Potential of Food Waste Chemistry for Biodegradable Plastics Production: Recent Advancements, Perspectives, and Life-Cycle Assessment

Trends in Food Science & Technology, Год журнала: 2024, Номер unknown, С. 104836 - 104836

Опубликована: Дек. 1, 2024

Язык: Английский

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Conversion of industrially relevant lignocellulosic biomass into monomers: By customization of cellulolytic enzyme cocktail using simplex-lattice mixture designing (SLMD)

Bioresource Technology Reports, Год журнала: 2024, Номер 27, С. 101891 - 101891

Опубликована: Июнь 20, 2024

Язык: Английский

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