Biochemical characterization of CYP505D6, a self-sufficient cytochrome P450 from the white-rot fungus Phanerochaete chrysosporium

Kiyota Sakai, Fumiko Matsuzaki, Lisa Wise, Yu Sakai, Sadanari Jindou, Hirofumi Ichinose, Naoki Takaya, Masashi Kato, Hiroyuki Wariishi, Motoyuki Shimizu

研究成果: ジャーナルへの寄稿記事

抄録

The activity of a self-sufficient cytochrome P450 enzyme, CYP505D6, from the lignin-degrading basidiomycete Phanerochaete chrysosporium was characterized. Recombinant CYP505D6 was produced in Escherichia coli and purified. In the presence of NADPH, CYP505D6 used a series of saturated fatty alcohols with C9-18 carbon chain lengths as the substrates. Hydroxylation occurred at the ω-1 to ω-6 positions of such substrates with C 9-15 carbon chain lengths, except for 1-dodecanol, which was hydroxylated at the ω-1 to ω-7 positions. Fatty acids were also substrates of CYP505D6. Based on the sequence alignment, the corresponding amino acid of Tyr51, which is located at the entrance to the active-site pocket in CYP102A1, was Val51 in CYP505D6. To understand the diverse hydroxylation mechanism, wild-type CYP505D6 and its V51Y variant and wild-type CYP102A1 and its Y51V variant were generated, and the products of their reaction with dodecanoic acid were analyzed. Compared with wild-type CYP505D6, its V51Y variant generated few products hydroxylated at the ω-4 to ω-6 positions. The products generated by wild-type CYP102A1 were hydroxylated at the ω-1 to ω-4 positions, whereas its Y51V variant generated ω-1 to ω-7 hydroxydodecanoic acids. These observations indicated that Val51 plays an important role in determining the regiospecificity of fatty acid hydroxylation, at least that at the ω-4 to ω-6 positions. Aromatic compounds, such as naphthalene and 1-naphthol, were also hydroxylated by CYP505D6. These findings highlight a unique broad substrate spectrum of CYP505D6, rendering it an attractive candidate enzyme for the biotechnological industry.

元の言語英語
記事番号e01091-18
ジャーナルApplied and environmental microbiology
84
発行部数22
DOI
出版物ステータス出版済み - 11 1 2018

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Phanerochaete
Phanerochaete chrysosporium
white-rot fungi
hydroxylation
Hydroxylation
cytochrome P-450
Cytochrome P-450 Enzyme System
cytochrome
lauric acid
Fungi
fungus
substrate
Dodecanol
Fatty Acids
Carbon
1-naphthol
Fatty Alcohols
fatty acids
fatty alcohols
Basidiomycota

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

これを引用

Biochemical characterization of CYP505D6, a self-sufficient cytochrome P450 from the white-rot fungus Phanerochaete chrysosporium. / Sakai, Kiyota; Matsuzaki, Fumiko; Wise, Lisa; Sakai, Yu; Jindou, Sadanari; Ichinose, Hirofumi; Takaya, Naoki; Kato, Masashi; Wariishi, Hiroyuki; Shimizu, Motoyuki.

:: Applied and environmental microbiology, 巻 84, 番号 22, e01091-18, 01.11.2018.

研究成果: ジャーナルへの寄稿記事

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abstract = "The activity of a self-sufficient cytochrome P450 enzyme, CYP505D6, from the lignin-degrading basidiomycete Phanerochaete chrysosporium was characterized. Recombinant CYP505D6 was produced in Escherichia coli and purified. In the presence of NADPH, CYP505D6 used a series of saturated fatty alcohols with C9-18 carbon chain lengths as the substrates. Hydroxylation occurred at the ω-1 to ω-6 positions of such substrates with C 9-15 carbon chain lengths, except for 1-dodecanol, which was hydroxylated at the ω-1 to ω-7 positions. Fatty acids were also substrates of CYP505D6. Based on the sequence alignment, the corresponding amino acid of Tyr51, which is located at the entrance to the active-site pocket in CYP102A1, was Val51 in CYP505D6. To understand the diverse hydroxylation mechanism, wild-type CYP505D6 and its V51Y variant and wild-type CYP102A1 and its Y51V variant were generated, and the products of their reaction with dodecanoic acid were analyzed. Compared with wild-type CYP505D6, its V51Y variant generated few products hydroxylated at the ω-4 to ω-6 positions. The products generated by wild-type CYP102A1 were hydroxylated at the ω-1 to ω-4 positions, whereas its Y51V variant generated ω-1 to ω-7 hydroxydodecanoic acids. These observations indicated that Val51 plays an important role in determining the regiospecificity of fatty acid hydroxylation, at least that at the ω-4 to ω-6 positions. Aromatic compounds, such as naphthalene and 1-naphthol, were also hydroxylated by CYP505D6. These findings highlight a unique broad substrate spectrum of CYP505D6, rendering it an attractive candidate enzyme for the biotechnological industry.",
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AU - Sakai, Kiyota

AU - Matsuzaki, Fumiko

AU - Wise, Lisa

AU - Sakai, Yu

AU - Jindou, Sadanari

AU - Ichinose, Hirofumi

AU - Takaya, Naoki

AU - Kato, Masashi

AU - Wariishi, Hiroyuki

AU - Shimizu, Motoyuki

PY - 2018/11/1

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