Mechanistic Study of Oxidoreductase AprQ Involved in Biosynthesis of Aminoglycoside Antibiotic Apramycin
Jinxiu Wang
State Key Laboratory of Cryospheric Science, Key Laboratory of Extreme Environmental Microbial Resources and Engineering, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou, Gansu, 730000 China
Department of Chemistry, Fudan University, Shanghai, 200433 China
University of Chinese Academy of Sciences, Beijing, 100049 China
Search for more papers by this authorSuze Ma
Department of Chemistry, Fudan University, Shanghai, 200433 China
Search for more papers by this authorWei Ding
State Key Laboratory of Microbial Metabolism, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240 China
Search for more papers by this authorCorresponding Author
Tuo Chen
State Key Laboratory of Cryospheric Science, Key Laboratory of Extreme Environmental Microbial Resources and Engineering, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou, Gansu, 730000 China
E-mail: [email protected]; [email protected]Search for more papers by this authorCorresponding Author
Qi Zhang
Department of Chemistry, Fudan University, Shanghai, 200433 China
E-mail: [email protected]; [email protected]Search for more papers by this authorJinxiu Wang
State Key Laboratory of Cryospheric Science, Key Laboratory of Extreme Environmental Microbial Resources and Engineering, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou, Gansu, 730000 China
Department of Chemistry, Fudan University, Shanghai, 200433 China
University of Chinese Academy of Sciences, Beijing, 100049 China
Search for more papers by this authorSuze Ma
Department of Chemistry, Fudan University, Shanghai, 200433 China
Search for more papers by this authorWei Ding
State Key Laboratory of Microbial Metabolism, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240 China
Search for more papers by this authorCorresponding Author
Tuo Chen
State Key Laboratory of Cryospheric Science, Key Laboratory of Extreme Environmental Microbial Resources and Engineering, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou, Gansu, 730000 China
E-mail: [email protected]; [email protected]Search for more papers by this authorCorresponding Author
Qi Zhang
Department of Chemistry, Fudan University, Shanghai, 200433 China
E-mail: [email protected]; [email protected]Search for more papers by this authorMain observation and conclusion
The aminoglycoside antibiotic apramycin contains a unique bicyclic octose moiety, and biosynthesis of this moiety involves an oxidoreductase AprQ. Unlike other known “Q” series proteins involved in aminoglycosides biosynthesis, AprQ does not work with an aminotransferase partner, and performs a four-electron oxidation that converts a CH2OH moiety to a carboxylate group. In this study, we report mechanistic investigation of AprQ. We showed AprQ contains a flavin mononucleotide (FMN) cofactor, which is different from other known Q series enzymes that contain a flavin adenine dinucleotide (FAD) cofactor. A series of biochemical assays showed that AprQ is not a monooxygenase but a flavoprotein oxidase. Although molecular O2 is strictly required for reaction turnover, four-electron oxidation can be achieved in the absence of O2 in single turnover condition. These findings establish the detailed catalytic mechanism of AprQ and expand the growing family of flavoprotein oxidases, an increasingly important class of biocatalysts.
Supporting Information
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Appendix S1: Supporting Information |
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