Atypical kinetics of cytochromes P450 catalysing 3'-hydroxylation of flavone from the white-rot fungus Phanerochaete chrysosporium

Noriyuki Kasai, Shinichi Ikushiro, Shinji Hirosue, Akira Arisawa, Hirofumi Ichinose, Yujirou Uchida, Hiroyuki Wariishi, Miho Ohta, Toshiyuki Sakaki

研究成果: ジャーナルへの寄稿記事

16 引用 (Scopus)

抄録

We cloned full-length cDNAs of 130 cytochrome P450s (P450s) derived from Phanerochaete chrysosporium and successfully expressed 70 isoforms in Saccharomyces cerevisiae. To elucidate substrate specificity of P. chrysosporium P450s, we examined various substrates including steroid hormones, several drugs, flavonoids and polycyclic aromatic hydrocarbons using the recombinant S. cerevisiae cells. Of these P450s, two CYPs designated as PcCYP50c and PcCYP142c with 14% identity in their amino acid sequences catalyse 3'-hydroxylation of flavone and O-deethylation of 7-ethoxycoumarin. Kinetic data of both enzymes on both reactions fitted not to the Michaelis-Menten equation but to Hill's equation with a coefficient of 2, suggesting that two substrates bind to the active site. Molecular modelling of PcCYP50c and a docking study of flavone to its active site supported this hypothesis. The enzymatic properties of PcCYP50c and PcCYP142c resemble mammalian drug-metabolizing P450s, suggesting that their physiological roles are metabolism of xenobiotics. It is noted that these unique P. chrysosporium P450s have a potential for the production of useful flavonoids.

元の言語英語
ページ(範囲)117-125
ページ数9
ジャーナルJournal of biochemistry
147
発行部数1
DOI
出版物ステータス出版済み - 1 1 2010

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flavone
Phanerochaete
Hydroxylation
Fungi
Cytochrome P-450 Enzyme System
Flavonoids
Yeast
Kinetics
Saccharomyces cerevisiae
Catalytic Domain
Substrates
Steroid hormones
Molecular modeling
Polycyclic Aromatic Hydrocarbons
Xenobiotics
Cytochromes
Substrate Specificity
Metabolism
Pharmaceutical Preparations
Amino Acid Sequence

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

これを引用

Atypical kinetics of cytochromes P450 catalysing 3'-hydroxylation of flavone from the white-rot fungus Phanerochaete chrysosporium. / Kasai, Noriyuki; Ikushiro, Shinichi; Hirosue, Shinji; Arisawa, Akira; Ichinose, Hirofumi; Uchida, Yujirou; Wariishi, Hiroyuki; Ohta, Miho; Sakaki, Toshiyuki.

:: Journal of biochemistry, 巻 147, 番号 1, 01.01.2010, p. 117-125.

研究成果: ジャーナルへの寄稿記事

Kasai, Noriyuki ; Ikushiro, Shinichi ; Hirosue, Shinji ; Arisawa, Akira ; Ichinose, Hirofumi ; Uchida, Yujirou ; Wariishi, Hiroyuki ; Ohta, Miho ; Sakaki, Toshiyuki. / Atypical kinetics of cytochromes P450 catalysing 3'-hydroxylation of flavone from the white-rot fungus Phanerochaete chrysosporium. :: Journal of biochemistry. 2010 ; 巻 147, 番号 1. pp. 117-125.
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abstract = "We cloned full-length cDNAs of 130 cytochrome P450s (P450s) derived from Phanerochaete chrysosporium and successfully expressed 70 isoforms in Saccharomyces cerevisiae. To elucidate substrate specificity of P. chrysosporium P450s, we examined various substrates including steroid hormones, several drugs, flavonoids and polycyclic aromatic hydrocarbons using the recombinant S. cerevisiae cells. Of these P450s, two CYPs designated as PcCYP50c and PcCYP142c with 14{\%} identity in their amino acid sequences catalyse 3'-hydroxylation of flavone and O-deethylation of 7-ethoxycoumarin. Kinetic data of both enzymes on both reactions fitted not to the Michaelis-Menten equation but to Hill's equation with a coefficient of 2, suggesting that two substrates bind to the active site. Molecular modelling of PcCYP50c and a docking study of flavone to its active site supported this hypothesis. The enzymatic properties of PcCYP50c and PcCYP142c resemble mammalian drug-metabolizing P450s, suggesting that their physiological roles are metabolism of xenobiotics. It is noted that these unique P. chrysosporium P450s have a potential for the production of useful flavonoids.",
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