文献类型：期刊文献

英文题名：Activity and stability improvement: structure-function insights into CotA from Bacillus subtilis

作者：He, Wei;Yiru;Wang;Ge, Xizhen;Li, Ying[1]

第一作者：He, Wei

通讯作者：Li, Y[1]

机构：[1]Beijing Union Univ, Coll Biochem Engieering, Beijing Key Lab Biomass Waste Resource Utilizat, Beijing 100023, Peoples R China; Beijing Union Univ, Coll Biochem Engn, Beijing Key Lab Bioact Subst & Funct Foods, Beijing 100023, Peoples R China

第一机构：北京联合大学应用文理学院|北京联合大学生物化学工程学院

通讯机构：[1]corresponding author), Beijing Union Univ, Coll Biochem Engieering, Beijing Key Lab Biomass Waste Resource Utilizat, Beijing 100023, Peoples R China.|[11417]北京联合大学;

年份：2025

卷号：318

外文期刊名：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES

收录：;EI(收录号：20252518635548);Scopus(收录号：2-s2.0-105008284675);WOS:【SCI-EXPANDED(收录号:WOS:001517352300022)】；

基金：This work is financially supported by the Project of Cultivation for Young Top-notch Talents of Beijing Municipal Institutions (BPHR202203209) .

语种：英文

外文关键词：CotA; Site-directed mutagenesis; MD simulation; Thermal stability; Catalytic efficiency

摘要：CotA, a bacterial multicopper oxidase, exhibits exceptional thermal stability, alkali resistance, and substrate versatility, making it valuable for industrial biocatalysis, environmental remediation, and dye degradation. Extensive research has enhanced its activity and stability through directed evolution, semi-rational design, and rational design, yet the underlying structural features remain largely unclear. In this study, we performed site-directed mutagenesis on CotA from Bacillus subtilis and analyzed its effects on stability. Among the CotA single and double mutants we constructed, the T377I/T418G variant exhibited a 6.12-fold increase in activity compared to the wild type and a 50 % improvement in thermal stability at 80 degrees C. To uncover the relationship between the enzyme activity, stability, and the spatial structure of the CotA mutants, we conducted molecular dynamics (MD) simulations to analyze the catalytic structural features. The results indicated that the increase in ABTS-specific enzyme activity was linked to an expanded binding pocket, while the increased thermal stability was attributed to a higher proportion of random coils in the secondary structure. Our study provides new insights into the catalytic activity and thermostability structural features of CotA, laying the groundwork for its applications in industrial and environmental fields.