Uncovering the cytochrome P450-catalyzed methylenedioxy bridge formation in streptovaricins biosynthesis.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
09 09 2020
09 09 2020
Historique:
received:
26
03
2020
accepted:
18
08
2020
entrez:
10
9
2020
pubmed:
11
9
2020
medline:
2
10
2020
Statut:
epublish
Résumé
Streptovaricin C is a naphthalenic ansamycin antibiotic structurally similar to rifamycins with potential anti-MRSA bioactivities. However, the formation mechanism of the most fascinating and bioactivity-related methylenedioxy bridge (MDB) moiety in streptovaricins is unclear. Based on genetic and biochemical evidences, we herein clarify that the P450 enzyme StvP2 catalyzes the MDB formation in streptovaricins, with an atypical substrate inhibition kinetics. Furthermore, X-ray crystal structures in complex with substrate and structure-based mutagenesis reveal the intrinsic details of the enzymatic reaction. The mechanism of MDB formation is proposed to be an intramolecular nucleophilic substitution resulting from the hydroxylation by the heme core and the keto-enol tautomerization via a crucial catalytic triad (Asp89-His92-Arg72) in StvP2. In addition, in vitro reconstitution uncovers that C6-O-methylation and C4-O-acetylation of streptovaricins are necessary prerequisites for the MDB formation. This work provides insight for the MDB formation and adds evidence in support of the functional versatility of P450 enzymes.
Identifiants
pubmed: 32908132
doi: 10.1038/s41467-020-18336-5
pii: 10.1038/s41467-020-18336-5
pmc: PMC7481197
doi:
Substances chimiques
Bacterial Proteins
0
Recombinant Proteins
0
Streptovaricin
1404-74-6
damavaricin C
58849-86-8
Cytochrome P-450 Enzyme System
9035-51-2
streptovaricin C
H41T05O2S2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
4501Références
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