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 - 1999 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Purification and characterization of 2-ethoxyphenol-induced cytochrome P450 from Corynebacterium sp. strain EP1
AU - Kawahara, Naoko
AU - Ikatsu, Hisayoshi
AU - Kawata, Hiroshige
AU - Miyoshi, Shin-ichi
AU - Tomochika, Ken ichi
AU - Sinoda, Sumio
PY - 1999
Y1 - 1999
N2 - 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.
AB - 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.
KW - 2-ethoxyphenol
KW - Biodegradation
KW - Corynebacterium sp.
KW - Cytochrome P450
KW - Enzyme purification
UR - http://www.scopus.com/inward/record.url?scp=0039700056&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0039700056&partnerID=8YFLogxK
M3 - Article
C2 - 10907420
AN - SCOPUS:0039700056
VL - 45
SP - 833
EP - 839
JO - Canadian Journal of Microbiology
JF - Canadian Journal of Microbiology
SN - 0008-4166
IS - 10
ER -