The enzymatic conversion of chitin to chitobiose is of significant interest due to its broad biological applications. However, the limited functional diversity and insufficient synergism of chitinase systems constrain catalytic efficiency and industrial applicability. In this study, we rationally reconstructed a cross-family chitinase cocktail by elucidating the synergistic mechanisms between GH18 and GH19 enzymes from Streptomyces sp. F-3. The three GH18 chitinases and one GH19 chitinase displayed complementary substrate specificity toward insoluble polysaccharides and oligosaccharides, suggesting potential for synergistic chitin degradation. Guided by the pattern of chitinase secretion, we designed the in vitro four-enzyme cocktail (FR48) based on the native stoichiometry at 48 h. FR48 exhibited remarkable performance, yielding 3814 mg/L chitobiose with 93.1% purity and achieving an 8.86-fold increase in reducing-sugar production over commercial chitinase under the tested conditions. This study provides a generalizable biomimetic strategy for designing cross-family multi-enzyme systems for recalcitrant biomass valorization in green biomanufacturing.
Sha Zhao, Fei-Yue Wen, Xiaomeng Sun et al.· Food Chemistry· 0 citations
A five-agent workflow to systematically identify divergent protein-membrane-interface sites across class A GPCRs and exploit them for selective allosteric ligand discovery is developed, establishing a scalable strategy for translating GPCR membrane-interface divergence into precise allosteric sites and testable subtype-selective ligand candidates.
Jing-Yi Zhu, Hengde Li, Min Xiao et al.· bioRxiv· 0 citations
The industrial application of enzyme catalysts is often constrained by the trade-off between thermostability and catalytic activity. Here, a region-focused engineering strategy was applied to a thermophilic GH10 xylanase to simultaneously improve both properties. The strategy integrates qProtein-guided hydrophobic cluster design for scaffold stabilization and dynamic loop analysis for active-site optimization. The resulting triple mutant A206S-N209D-F130L exhibited substantially improved thermostability, with a 5.79 °C increase in melting temperature and an 18.8-fold extension of the half-life at 60 °C. Its optimum temperature increased from 60 to 70 °C, accompanied by a 129.4% enhancement in catalytic activity at 70 °C relatively to the wild type. Molecular dynamics simulations indicated that these mutations reshape the conformational energy landscape by stabilizing hydrophobic packing and modulating loop dynamics. This study provides a generalizable framework for simultaneously improving enzyme stability and catalytic performance.
Zhaoran Li, Zhixin Dou, Sha Zhao et al.· Journal of Agricultural and...· 0 citations
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