The Diiron Monooxygenase CmlA from Chloramphenicol Biosynthesis Allows Reconstitution of β‑Hydroxylation during Glycopeptide Antibiotic Biosynthesis

β-Hydroxylation plays an important role in the nonribosomal peptide biosynthesis of many important natural products, including bleomycin, chloramphenicol, and the glycopeptide antibiotics (GPAs). Various oxidative enzymes have been implicated in such a process, with the mechanism of incorporation va...

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Bibliographic Details
Published in:ACS chemical biology 2019-12, Vol.14 (12), p.2932-2941
Main Authors: Kaniusaite, Milda, Goode, Robert J. A, Schittenhelm, Ralf B, Makris, Thomas M, Cryle, Max J
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Anti-Bacterial Agents - biosynthesis
Chloramphenicol - biosynthesis
Glycopeptides - biosynthesis
Hydroxylation
Iron - metabolism
Mixed Function Oxygenases - chemistry
Mixed Function Oxygenases - metabolism
Substrate Specificity
Teicoplanin - biosynthesis
Tyrosine - metabolism
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Summary:β-Hydroxylation plays an important role in the nonribosomal peptide biosynthesis of many important natural products, including bleomycin, chloramphenicol, and the glycopeptide antibiotics (GPAs). Various oxidative enzymes have been implicated in such a process, with the mechanism of incorporation varying from installation of hydroxyl groups in amino acid precursors prior to adenylation to direct amino acid oxidation during peptide assembly. In this work, we demonstrate the in vitro utility and scope of the unusual nonheme diiron monooxygenase CmlA from chloramphenicol biosynthesis for the β-hydroxylation of a diverse range of carrier protein bound substrates by adapting this enzyme as a non-native trans-acting enzyme within NRPS-mediated GPA biosynthesis. The results from our study show that CmlA has a broad substrate specificity for modified phenylalanine/tyrosine residues as substrates and can be used in a practical strategy to functionally cross complement compatible NRPS biosynthesis pathways in vitro.
ISSN:1554-8929
1554-8937
DOI:10.1021/acschembio.9b00862