Thermostability Improvement of the Chitinase from Bacillus circulans for Efficient Chitin Oligosaccharide Production via Computational Design

Biomolecules, Год журнала: 2025, Номер 15(3), С. 330 - 330

Опубликована: Фев. 24, 2025

The chitinase A1 from Bacillus circulans WL-12 (BcChiA1) exhibits promising potential for producing chitin oligosaccharides (CHOs), while its application is limited by poor thermal stability. In this study, a set of thermostable variants were obtained modifying BcChiA1 using comprehensive strategy based on computer-aided design. A combination five beneficial single-point mutations (S67G/K177R/A220V/N257Y/N271E) to generated markedly improved variant, Mu5. Mu5 exhibited half-life 295 min at 60 °C, which was 59 times higher than that BcChiA1. Furthermore, reused conversion, releasing 86.14 ± 3.73 mM CHOs after reaction cycles. Molecular dynamics simulation and structural analysis revealed these enhancements driven increased rigidity compactness, resulting in protein conformation less prone denaturation. This combined approach through computational design yielded potentially facilitating production economical way.

Язык: Английский

FADS-Based Directed Evolution of a Robust Bst DNA Polymerase Adapting High-Temperature Loop-Mediated Isothermal Amplification (HT-LAMP)

ACS Catalysis, Год журнала: 2025, Номер unknown, С. 5391 - 5402

Опубликована: Март 18, 2025

Язык: Английский