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

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

Faculty of Pharmaceutical Sciences

Research output: Contribution to journal › Article › peer-review

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 language	English
Pages (from-to)	833-839
Number of pages	7
Journal	Canadian journal of microbiology
Volume	45
Issue number	10
Publication status	Published - Dec 1 1999

Keywords

2-ethoxyphenol
Biodegradation
Corynebacterium sp.
Cytochrome P450
Enzyme purification

ASJC Scopus subject areas

Microbiology
Immunology
Applied Microbiology and Biotechnology
Molecular Biology
Genetics

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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, 01.12.1999, p. 833-839.

Research output: Contribution to journal › Article › peer-review

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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.",

keywords = "2-ethoxyphenol, Biodegradation, Corynebacterium sp., Cytochrome P450, Enzyme purification",

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AU - Miyoshi, Shin Ichi

AU - Tomochika, Ken ichi

AU - Sinoda, Sumio

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