In Vitro Reconstitution of OxyA Enzymatic Activity Clarifies Late Steps in Vancomycin Biosynthesis.

TitleIn Vitro Reconstitution of OxyA Enzymatic Activity Clarifies Late Steps in Vancomycin Biosynthesis.
Publication TypeJournal Article
Year of Publication2017
AuthorsForneris, CC, Ozturk, S, Gibson, MI, Sorensen, EJ, Seyedsayamdost, MR
JournalACS Chem Biol
Volume12
Issue9
Pagination2248-2253
Date Published2017 09 15
ISSN1554-8937
KeywordsActinomycetales, Anti-Bacterial Agents, Biosynthetic Pathways, Cytochrome P-450 Enzyme System, Halogenation, Substrate Specificity, Vancomycin
Abstract

Studies on the biosynthesis of glycopeptide antibiotics have provided many insights into the strategies that Nature employs to build architecturally strained molecules. A key structural feature of vancomycin, the founding member of this class, is a set of three aromatic cross-links that are introduced via yet unknown mechanisms. Previous reports have identified three cytochrome P450 enzymes involved in this process and demonstrated enzymatic activity for OxyB, which installs the first aromatic cross-link. However, the activities of the remaining two P450 enzymes have not been recapitulated. Herein, we show that OxyA generates the second bis-aryl ether bond in vancomycin and that it exhibits strict substrate specificity toward the chlorinated, OxyB-cross-linked product. No OxyA product is detected with the unchlorinated substrate. Together with previous results, these data suggest that chlorination occurs after OxyB- but before OxyA-catalyzed cross-link formation. Our results have important implications for the chemo-enzymatic synthesis of vancomycin and its analogs.

DOI10.1021/acschembio.7b00456
Alternate JournalACS Chem. Biol.
PubMed ID28696669
PubMed Central IDPMC5617736
Grant ListDP2 AI124786 / AI / NIAID NIH HHS / United States
R01 GM065483 / GM / NIGMS NIH HHS / United States