Molecular anatomy of 2-oxoacid : ferredoxin oxidoreductase from hyperthermophilic archaea

2-含氧酸的分子解剖学：来自超嗜热古菌的铁氧还蛋白氧化还原酶

• 批准号: 09660074
• 负责人: WAKAGI Takayoshi
• 金额: $ 2.11万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09660074/
• 关键词: archaea; thermophile; oxidoreductase; ferredoxin; 2-オキソ酸; 耐熱性酵素; 遺伝子発現

2-Oxoacid : ferredoxin oxidoreductase (OFOR) catalyzes oxidative decarboxylation of 2-oxoacids, using ferredoxin as an electron acceptor. The OFOR is found among all archaea, certain eubacteria, and certain eukaryotes. The structure-function relationship of OFOR is not clearly elucidated. This is partly because of the difficulty in constructing gene expression system that produces stable enzyme, or of the intrinsic oxygen-sensitivity of most OFORS from anaerobic organisms. In the present study, we first constructed the OFOR gene expression system from Sulfolobus sp. strain 7, compared the characteristics of recombinant enzyme with natural one. The two OFORs were indistinguishable by the criteria so far investigated. Based on the gene expression system, a series of mutants that is implicated as catalytic sites were designed. Some of them lost activity, and the other were found to change its substrate specificity.

2-氧化物：使用铁蛋白作为电子受体，使用铁蛋白氧化剂氧化剂（OFOR）催化2-氧化物的氧化脱羧。在所有古细菌，某些Eubacteria和某些真核生物中都发现了Ofor。 OFOR的结构功能关系尚未清楚地阐明。这部分是因为难以构建产生稳定酶的基因表达系统，或者是大多数厌氧生物的固有氧气敏感性。在本研究中，我们首先从Sulfolobus sp。构建了OFOR基因表达系统。菌株7，将重组酶与天然酶的特征进行了比较。到目前为止，这两个官方的标准无法区分。基于基因表达系统，设计了一系列与催化位点有关的突变体。他们中的一些人失去了活动，另一些人被发现改变了其底物特异性。

项目成果

福田英理子、松澤洋、若木高善: "好酸好熱性古細菌Sulfololbus sp.strain7の2-オキソ酸:フェレドキシン酸化還元酵素の構造と機能"日本農芸化学会誌. 73. 217 (1999)

Eriko Fukuda、Hiroshi Matsuzawa、Takayoshi Wakagi：“嗜酸古菌 Sulfololbus sp. 菌株 2-含氧酸：铁氧还蛋白氧化还原酶的结构和功能”日本农业化学学会杂志 73. 217 (1999)。

Kojoh, K., Matsuzawa, H., Wakagi, T.: "Zinc and an N-terminal extra stretch of the ferredoxin from a thermoacidophilic archaeon stabilize the molecule at high temperature"Eur J Biochem.. 264. 85-91 (1999)

Kojoh, K.、Matsuzawa, H.、Wakagi, T.：“锌和来自嗜热嗜酸古菌的铁氧还蛋白 N 端额外延伸在高温下稳定分子”Eur J Biochem.. 264. 85-91 (1999

Wakagi, T., and Ihara, Y.: "Electron transport system and ATPase""Biology of Archaea" (ed. Koga, Y. and Kamekura, M.). 217-238 (1998)

Wakagi, T. 和 Ihara, Y.：“电子传输系统和 ATP 酶”“古细菌生物学”（Koga, Y. 和 Kamekura, M. 编辑）。

伏信進矢ほか: "Crystallographic and mutational analyses of an extremely acidophilic and acid-stable xylanase biased distribution of acidic residues and importance of Asp37 for catalysis at low pH." Protein Engineering.11(12). 1121-1128 (1998)

Shinya Fushinobu 等人：“酸性残基的极端嗜酸和酸稳定木聚糖酶偏向分布的晶体学和突变分析以及 Asp37 在低 pH 条件下催化的重要性。11(12) (1998)。”

Nishihara, M. and Wakagi, T.: "Central metabolism""Biology of Archaea" (ed. Koga, Y. and Kamekura, M.). 217-238 (1998)

Nishihara, M. 和 Wakagi, T.：“中枢代谢”“古细菌生物学”（Koga, Y. 和 Kamekura, M. 编辑）。