Purification and characterization of 2-ethoxyphenol-induced cytochrome P450 from Corynebacterium sp. strain EP1

Naoko Kawahara, Hisayoshi Ikatsu, Hiroshige Kawata, Shin-ichi Miyoshi, Ken ichi Tomochika, Sumio Sinoda

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)833-839
Number of pages7
JournalCanadian Journal of Microbiology
Volume45
Issue number10
Publication statusPublished - 1999
Externally publishedYes

Fingerprint

Corynebacterium
Cytochrome P-450 Enzyme System
Purification
Carbon Monoxide
Ferredoxin-NADP Reductase
Dealkylation
Guaiacol
Spinacia oleracea
Carbon Tetrachloride
Toluene
Benzene
Electrophoresis
NADP
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Assays
Molecular Weight
Molecular weight
guaethol
Substrates

Keywords

  • 2-ethoxyphenol
  • Biodegradation
  • Corynebacterium sp.
  • Cytochrome P450
  • Enzyme purification

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Immunology
  • Microbiology

Cite this

Purification and characterization of 2-ethoxyphenol-induced cytochrome P450 from Corynebacterium sp. strain EP1. / Kawahara, Naoko; Ikatsu, Hisayoshi; Kawata, Hiroshige; Miyoshi, Shin-ichi; Tomochika, Ken ichi; Sinoda, Sumio.

In: Canadian Journal of Microbiology, Vol. 45, No. 10, 1999, p. 833-839.

Research output: Contribution to journalArticle

Kawahara, Naoko ; Ikatsu, Hisayoshi ; Kawata, Hiroshige ; Miyoshi, Shin-ichi ; Tomochika, Ken ichi ; Sinoda, Sumio. / Purification and characterization of 2-ethoxyphenol-induced cytochrome P450 from Corynebacterium sp. strain EP1. In: Canadian Journal of Microbiology. 1999 ; Vol. 45, No. 10. pp. 833-839.
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