Molecular design of hydrogen peroxide-dependent P450s
| Project/Area Number |
17H03087
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bio-related chemistry
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| Research Institution | Nagoya University |
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Principal Investigator |
Watanabe Yoshihito 名古屋大学, 物質科学国際研究センター, 名誉教授 (10201245)
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| Project Period (FY) |
2017-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000)
Fiscal Year 2019: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
Fiscal Year 2018: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
Fiscal Year 2017: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
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| Keywords | 酵素 / シトクロムP450 / ヘム / 過酸化水素 / 水酸化反応 / カタラーゼ / 結晶構造解析 / 変異導入 / 触媒 / 鉄ポルフィリン錯体 / 酸化反応 / 不均化反応 / 酸素 |
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| Outline of Final Research Achievements |
CYP152A1 (P450BSβ) and CYP152B1 (P450SPα) can generate the active species utilizing H2O2 as an oxidant and catalyze the α- or β-selective hydroxylation of long-alkyl-chain fatty acids. The substrate specificity of CYP152 family enzymes is very high. The crystal structure of P450BSβ and P450SPα indicate that the carboxylate group of palmitic acid interacting with the arginine located at the I helix (the distal side of the heme) is crucial for the generation of compound I. Owing to this unique mechanism, CYP152 family enzymes never oxidize substrate other than long-alkyl-chain fatty acid. We have demonstrated that the R242K mutant of P450BSβ and the P450SPα with corresponding mutation can catalyze non-native substrate oxidations. We have also succeeded in crystal structure analysis of the R241K mutant of P450SPα. We expect that a variety of peroxygenases based on P450s can be constructed by one-point mutagenesis.
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| Academic Significance and Societal Importance of the Research Achievements |
シトクロムP450は、高難度酸化反応が可能な有用な酸化酵素の一つですが、酸化反応を行うためには、高価な試薬であるNAD(P)Hが必要になります。安価な過酸化水素を利用することが可能なシトクロムP450もありますが、対象とする基質の選択性が高く、長鎖脂肪酸の水酸化しかできないと考えられてきました。本研究では、過酸化水素を利用可能なP450の一つのアミノ酸を違うアミノ酸に置換することで、様々な基質を水酸化できるように改変することに成功しました。
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Report
(4 results)
Research Products
(17 results)
