5y1i
The crystal structure of GfsF
Structural highlights
FunctionGFSF_STRHA Involved in the synthesis of the 16-membered macrolide antibiotics FD-891 and FD-892 (PubMed:20589823, PubMed:26630077, PubMed:28869713). Consecutively catalyzes epoxidation of C8-C9 and then hydroxylation at C10 to convert 25-O-methyl-FD-892 to FD-891 (PubMed:20589823). Consecutively catalyzes epoxidation of C8-C9 and then hydroxylation at C10 to convert 8,9-epoxy-FD-892 to 25-O-demethyl-FD-891 as well as converting 25-oxo-FD-892 to 8,9-epoxy-25-oxo-FD-892 and 8,9-epoxy-10-hydroxy-25-oxo-FD-892 (PubMed:26630077). In vitro is furnished with P.putida putidaredoxin and putidaredoxin reductase to provide the required two-electron reduction (PubMed:20589823, PubMed:26630077, PubMed:28869713).[1] [2] [3] Publication Abstract from PubMedGfsF is a multifunctional P450 monooxygenase that catalyzes the epoxidation and subsequent hydroxylation in the biosynthesis of macrolide polyketide FD-891. Here, we describe the biochemical and structural analysis of GfsF. To obtain the structural basis of a dual functional reaction, we determined the crystal structure of ligand-free GfsF, which revealed GfsF to have a predominantly hydrophobic substrate binding pocket. The docking models in conjunction with the results of the enzymatic assay with substrate analogs as well as site-directed mutagenesis suggested two distinct substrate binding modes for epoxidation and hydroxylation reactions, which explained how GfsF regulates the order of two oxidative reactions. These findings provide new insights into the reaction mechanism of multifunctional P450 monooxygenases. Substrate recognition by a dual functional P450 monooxygenase GfsF involved in FD-891 biosynthesis.,Eguchi T, Miyanaga A, Takayanagi R, Furuya T, Kawamata A, Itagaki T, Iwabuchi Y, Kanoh N, Kudo F Chembiochem. 2017 Sep 4. doi: 10.1002/cbic.201700429. PMID:28869713[4] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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