Structural characterization and fatty acid epoxidation of CYP184A1 from Streptomyces avermitilis

Arch Biochem Biophys. 2022 Sep 30:727:109338. doi: 10.1016/j.abb.2022.109338. Epub 2022 Jun 30.

Abstract

The genome of Streptomyces avermitilis contains 33 cytochrome P450 genes. Among the P450 gene products of S. avermitilis, we characterized the biochemical function and structural aspects of CYP184A1. Ultra-performance liquid chromatography-tandem mass spectrometry analysis showed that CYP184A1 induced an epoxidation reaction to produce 9,10-epoxystearic acid. Steady-state kinetic analysis yielded a kcat value of 0.0067 min-1 and a Km value 10 μM. The analysis of its crystal structures illustrated that the overall CYP184A1 structure adopts the canonical scaffold of cytochrome P450 and possesses a narrow and deep substrate pocket architecture that is required for binding to linear chain fatty acids. In the structure of the CYP184A1 oleic acid complex (CYP184A1-OA), C9-C10 of oleic acid was bound to heme for the productive epoxidation reaction. This study elucidates the roles of P450 enzymes in the oxidative metabolism of fatty acids in Streptomyces species.

Keywords: CYP; CYP184A1; Fatty acid; Oleic acid; P450; Streptomyces avermitilis; X-ray crystal structure.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cytochrome P-450 Enzyme System / chemistry
  • Fatty Acids* / metabolism
  • Kinetics
  • Oleic Acids / metabolism
  • Streptomyces*

Substances

  • Fatty Acids
  • Oleic Acids
  • Cytochrome P-450 Enzyme System

Supplementary concepts

  • Streptomyces avermitilis