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Article Abstract
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 with controlled degrees of polymerization. To enhance the industrial applicability of CsnMY002, molecular dynamics (MD) simulations were applied to investigate the structure-property relationship. Guided by the simulation results, the beneficial mutants were screened through a synergistic strategy using a residue-folding free energy calculation and consensus sequence analysis. Iterative combinations constructed the mutant Mut6 (A49G/K70A/S84A/N89G/D199R/N221G) with significantly improved thermal stability, which had a half-life ( value) at 55 °C and 75 °C that was 1.80 and 1.62 times higher than that of the wild type, respectively. A highly active mutant, Mut2, was created, exhibiting a 1.52 times catalytic efficiency of the wild type. An MD simulation analysis of the mutants suggested that the improved enzymatic properties were highly correlated with changes in the dynamic behaviours of the enzyme structure. This study generated more suitable CsnMY002 variants for COS production and provided a comprehensive strategy for the optimization of other industrial enzymes with application potential.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11855910 | PMC |
| http://dx.doi.org/10.3390/ijms26041588 | DOI Listing |
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