Enhancing the activity of disulfide-bond-containing proteins via promoting disulfide bond formation in Bacillus licheniformis

作者全名："Wang, Shiyi; Zhao, Yiwen; Mao, Shufen; Zhu, Jiang; Zhan, Yangyang; Cai, Dongbo; Ma, Xin; Wang, Dong; Chen, Shouwen"

作者地址："[Wang, Shiyi; Zhao, Yiwen; Mao, Shufen; Zhu, Jiang; Zhan, Yangyang; Cai, Dongbo; Ma, Xin; Chen, Shouwen] Hubei Univ, Coll Life Sci, Environm Microbial Technol Ctr Hubei Prov, State Key Lab Biocatalysis & Enzyme Engn, Wuhan 430062, Peoples R China; [Wang, Dong] Chongqing Med Univ, Sch Basic Med Sci, Chongqing, Peoples R China; [Wang, Dong; Chen, Shouwen] 368 Youyi Ave, Wuhan 430062, Hubei, Peoples R China"

通信作者："Chen, SW (通讯作者)，Hubei Univ, Coll Life Sci, Environm Microbial Technol Ctr Hubei Prov, State Key Lab Biocatalysis & Enzyme Engn, Wuhan 430062, Peoples R China.; Wang, D; Chen, SW (通讯作者)，368 Youyi Ave, Wuhan 430062, Hubei, Peoples R China."

来源：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES

ESI学科分类：BIOLOGY & BIOCHEMISTRY

WOS号：WOS:000936683600001

JCR分区：Q1

影响因子：7.7

年份：2023

卷号：233

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：Disulfide bonds; Disulfide -bond -containing proteins; Thiol-disulfide isomerase; Thiol-disulfide oxidoreductases; ?-Aminobutyric acid; Bacillus licheniformis

摘要："Disulfide bonds in proteins have strongly influence on the folding efficiency by constraining the conformational space. The inefficient disulfide bond formation of proteins is the main limiting factor of enzyme activity and stability. This study aimed to increase the activity of disulfide-bond-containing proteins via promoting disulfide bonds formation in Bacillus licheniformis. Initially, the glutamate decarboxylase GAD from Escherichia coli was selected as the model protein and introduced into the B. licheniformis. Then, the disulfide isomerase and oxidoreductase from different sources were excavated and overexpressed successively to improve the catalytic efficiency of GAD. The final engineered B. licheniformis showed significantly improved GAD specific activity (from 10.4 U/mg to 80.0 U/mg), which also presented perfect adaptability for other disulfide-bond-containing proteins, for instance, UDP-glucosyltransferase from Arabidopsis thaliana. Taken together, our work demon-strated that the activity of GAD in B. licheniformis was regulated by the disulfide bonds formation status and provided a promising platform for the expression of disulfide-bond-containing proteins."

基金机构：National Natural Science Foundation of China [31900026]; National Key Research and Development Program of China [2021YFC2100202]

基金资助正文：This work was supported by the National Natural Science Foundation of China (31900026) and the National Key Research and Development Program of China (2021YFC2100202) .