Coupling efficiency in light-driven hybrid P450BM3 and CYP119 enzymes

Arch Biochem Biophys. 2019 Sep 15:672:108077. doi: 10.1016/j.abb.2019.108077. Epub 2019 Aug 16.

Abstract

The light-driven hybrid P450 enzyme approach utilizing the photochemical properties of a covalently attached Ru(II)-diimine photosensitizer was extended to the archaeal Sulfolobus acidocaldarius CYP119 enzyme leading to high photocatalytic activity in the hydroxylation of the chromogenic substrate, 11-nitrophenoxyundecanoic acid. The determined kcat was greater than those reported with various natural redox partners. In addition, the sacrificial electron donor, diethyldithiocarbamate, used in the photocatalytic reaction is shown to play a dual role. It acts as an efficient quencher of the Ru(II) excited state leading to a highly reducing species necessary to inject electrons into the heme. It is also known for its antioxidant properties and is shown herein to be a useful probe to determine coupling efficiency in the light-driven hybrid enzymes.

Keywords: Bacterial P450 enzymes; Light-driven process; Reactive oxygen species; Uncoupling pathway.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Archaeal Proteins / chemistry*
  • Archaeal Proteins / genetics
  • Archaeal Proteins / radiation effects
  • Biocatalysis / radiation effects
  • Coordination Complexes / chemistry
  • Coordination Complexes / radiation effects
  • Cytochrome P-450 Enzyme System / chemistry*
  • Cytochrome P-450 Enzyme System / genetics
  • Cytochrome P-450 Enzyme System / radiation effects
  • Ditiocarb / chemistry
  • Heme / chemistry
  • Kinetics
  • Light
  • Mutation
  • Photosensitizing Agents / chemistry
  • Photosensitizing Agents / radiation effects
  • Ruthenium / chemistry
  • Sulfolobus acidocaldarius / enzymology

Substances

  • Archaeal Proteins
  • Coordination Complexes
  • Photosensitizing Agents
  • Heme
  • Ruthenium
  • Cytochrome P-450 Enzyme System
  • Ditiocarb