Mechanistic insights into dideoxygenation in gentamicin biosynthesis
Mechanistic insights into dideoxygenation in gentamicin biosynthesis
Abstract Gentamicin is an important aminoglycoside antibiotic used for treatment of infections caused by Gram-negative bacteria. Although most of the biosynthetic pathway of gentamicin has been elucidated, a remaining intriguing question is how the intermediates JI-20A and JI-20B undergo a dideoxygenation to form gentamicin C complex. Here we show that the dideoxygenation process starts with GenP-catalyzed phosphorylation of JI-20A and JI-20Ba. The phosphorylated products are converted to C1a and C2a by concerted actions of two PLP (pyridoxal 5’-phosphate)-dependent enzymes: elimination of water and then phosphate by GenB3 and double bond migration by GenB4. Each of these reactions liberates an imine which hydrolyses to a ketone or aldehyde and is then re-aminated by GenB3 using an amino donor. Crystal structures of GenB3 and GenB4 have guided site-directed mutagenesis to reveal crucial residues for the enzymes’ functions. We propose catalytic mechanisms for GenB3 and GenB4, which shed new light on the already unrivalled catalytic versatility of PLP-dependent enzymes.
Huang Fanglu、Guo Junhong、Huang Chuan、Jian Xinyun、Zhou Jiahai、Deng Zixin、Leeper Finian J.、Leadlay Peter F.、Bury Priscila dos Santos、Li Yuan、Reva Anna、Sun Guo、Dias Marcio V. B.、Li Sicong、Sun Yuhui
Department of Biochemistry, University of CambridgeKey Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan UniversityKey Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan UniversityKey Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan UniversityCAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institute of Advanced Technology, Chinese Academy of SciencesKey Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan UniversityDepartment of Chemistry, University of CambridgeDepartment of Biochemistry, University of CambridgeDepartment of Microbiology, Institute of Biomedical Science, University of S?o PauloKey Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan UniversityDepartment of Biochemistry, University of CambridgeKey Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan UniversityDepartment of Microbiology, Institute of Biomedical Science, University of S?o Paulo||Department of Chemistry, University of WarwickKey Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan UniversityKey Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan University
药学生物化学分子生物学
Huang Fanglu,Guo Junhong,Huang Chuan,Jian Xinyun,Zhou Jiahai,Deng Zixin,Leeper Finian J.,Leadlay Peter F.,Bury Priscila dos Santos,Li Yuan,Reva Anna,Sun Guo,Dias Marcio V. B.,Li Sicong,Sun Yuhui.Mechanistic insights into dideoxygenation in gentamicin biosynthesis[EB/OL].(2025-03-28)[2025-04-27].https://www.biorxiv.org/content/10.1101/2021.05.12.443773.点此复制
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