title: Differences in PLP-Dependent Cysteinyl Processing Lead to Diverse S-Functionalization of Lincosamide Antibiotics.
abstract: Pyridoxal-5'-phosphate (PLP)-dependent proteins constitute one of the largest and most important families of enzymes in living organisms. These proteins participate in numerous biochemical processes, many of which have not been characterized, and transform substrates containing an amino group through various reactions that share aldimine as a common intermediate. Herein, we report that the PLP-dependent enzymes CcbF and LmbF, which are highly related in phylogenesis, process cysteine S-conjugated intermediates in different ways and associate with individual downstream enzyme(s) toward distinct S-functionalization of the lincosamide antibiotics celesticetin and lincomycin A. CcbF catalyzes an unusual conversion that involves decarboxylation-coupled oxidative deamination of the cysteinyl group during the formation of a two-carbon alcohol linker, whereas LmbF is responsible for β-elimination, followed by S-methylation to produce a methylmercapto group. The two tailoring routes are variable and exchangeable with each other, allowing for in vitro combinatorial biosynthesis of a number of hybrid lincosamide antibiotics, including the natural product Bu-2545. These findings demonstrate the wide diversity of PLP chemistry in enzymatic catalysis and its promising applicability in creation of new molecules.
authors.affiliation: State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, University of Chinese Academy of Sciences , 345 Lingling Road, Shanghai 200032, China.
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, University of Chinese Academy of Sciences , 345 Lingling Road, Shanghai 200032, China.|State Key Laboratory of Microbial Metabolism, School of Life Science & Biotechnology, Shanghai Jiao Tong University , 800 Dongchuan Road, Shanghai 200240, China.
authors.forename: Qunfei
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: Huzhou Center of Bio-Synthetic Innovation , 1366 Hongfeng Road, Huzhou 313000, China.
authors.forename: Qinglin
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, University of Chinese Academy of Sciences , 345 Lingling Road, Shanghai 200032, China.|State Key Laboratory of Microbial Metabolism, School of Life Science & Biotechnology, Shanghai Jiao Tong University , 800 Dongchuan Road, Shanghai 200240, China.|Huzhou Center of Bio-Synthetic Innovation , 1366 Hongfeng Road, Huzhou 313000, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Liu
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry*; D002384:Catalysis; D003545:Cysteine / Q000737:chemistry*; D055231:Lincosamides / Q000737:chemistry*; D011732:Pyridoxal Phosphate / Q000737:chemistry*
chemical_list: D000900:Anti-Bacterial Agents; D055231:Lincosamides; D011732:Pyridoxal Phosphate; D003545:Cysteine