文献类型：期刊文献

英文题名：ADP-ribose is a competitive inhibitor of methanol dehydrogenases from Bacillus methanolicus

作者：Ma, Bao-Di[1,2,3];Li, Jia-Yi[1,2,4];Xu, Jian-He[4];Yu, Tao[5];Kong, Xu-Dong[1,2]

机构：[1]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, State Key Lab Microbial Metab, Shanghai, Peoples R China;[2]Shanghai Jiao Tong Univ, Zhangjiang Inst Adv Study, Shanghai, Peoples R China;[3]Shanghai Inst Technol, Sch Chem & Environm Engn, Shanghai, Peoples R China;[4]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg, State Key Lab Bioreactor Engn, Shanghai, Peoples R China;[5]Chinese Acad Sci, Shenzhen Inst Synthet Biol,Shenzhen Inst Adv Techn, Shenzhen Key Lab Intelligent Microbial Mfg Med, Ctr Synthet Biochem,Key Lab Quantitat Synthet Biol, Shenzhen, Peoples R China

年份：2025

卷号：301

期号：9

外文期刊名：JOURNAL OF BIOLOGICAL CHEMISTRY

收录：;EI(收录号：20253719138946);WOS:【SCI-EXPANDED(收录号:WOS:001572015900003)】；

基金：This work was financially supported by the Ministry of Science and Technology of the People's Republic of China (National Key R&D Program of China: 2021YFA0911000) and Shanghai Municipal Science and Technology Major Project.

语种：英文

外文关键词：Bacteriology - Biochemistry - Chemical activation - Crystallography - Hydrolases - Hydrolysis - Metabolic engineering - Metabolism - Methanol

摘要：Methanol dehydrogenase (MDH), a representative of Type III alcohol dehydrogenases (ADHs), plays a pivotal role in methanol assimilation pathways, making it a key enzyme for the biosynthesis of chemicals and fuels from one-carbon feedstocks. An activator protein belonging to the Nudix hydrolase family, ACT, was found to increase the activity of MDH by 40-fold. Despite the widespread observation of this in vitro activation phenomenon in pairs of type III alcohol dehydrogenases and Nudix hydrolases, the mechanistic details have remained unresolved for decades. Here, we uncover a regulation mechanism in which MDH activation arises from the hydrolytic removal of ADP-ribose (ADPR), a potent inhibitor derived from NAD+ degradation, by the ADPRase activity of ACT. This discovery challenges the previously proposed 'activation' models, revealing that ACT-mediated ADPR clearance disinhibits MDH rather than directly enhancing catalysis. By combining crystallographic analysis, kinetics, and inhibition assays, we demonstrate that ADPR inhibits MDHs with submicromolar Ki values, highlighting its potential regulatory role in metabolic networks. Our findings redefine the widespread 'activation' of type III ADHs, providing valuable insights into alcohol metabolism and new directions for engineering synthetic methanol utilization pathways.