A soluble cytochrome P450 (P450(EP1A)) induced by 2-ethoxyphenol was purified to apparent homogeneity from Corynebacterium sp. strain EP1. The P450(EP1A) showed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a molecular weight of about 45 kDa. The CO-reduced difference spectra of P450(EP1A) had a Soret maximum at 447.6 nm. The substrate difference spectra with 2-ethoxyphenol showed an absorption maximum at 394.0 nm. The purified P450(EP1A) degraded 2-ethoxyphenol in an assay system composed of spinach ferredoxin-NADP+ oxidoreductase and NADPH. The reaction activity decreased to 1.4% of its original activity by addition of CO. The existence of catechol in the reaction mixture was confirmed after the metabolic reaction, indicating that P450(EP1A) catalyzes O-dealkylation of 2-ethoxyphenol. In addition to 2-ethoxyphenol, the P450(EP1A) metabolized 2-methoxyphenol, 1,1,1-trichloroethane, carbon tetrachloride, benzene, and toluene.
|Number of pages||7|
|Journal||Canadian journal of microbiology|
|Publication status||Published - Dec 1 1999|
- Corynebacterium sp.
- Cytochrome P450
- Enzyme purification
ASJC Scopus subject areas
- Applied Microbiology and Biotechnology
- Molecular Biology