Identification and characterization of novel systems for oxygen metabolism and anti-oxidative stress protection in anaerobes

厌氧菌氧代谢和抗氧化应激保护新系统的鉴定和表征

• 批准号: 20580086
• 负责人: KAWASAKI Shinji
• 金额: $ 3.08万
• 依托单位: Tokyo University of Agriculture
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20580086/
• 关键词: 嫌気性菌; 抗酸化酵素; 活性酸素; 酸素毒性; Clostridium; Bifidobacterium; 腸内細菌; 嫌気性細菌; 活性酸素消去; ビフィズス菌; プロバイオティクス; 酸化ストレス; 酸素

Our purpose is to know the mechanism of O_2 sensitivity of the obligatory anaerobes Clostridium and Bifidomacterium. Clostridium acetobutylicum, which is well known as an efficient acetone-butanol fermenting bacteria, senses the contamination of O_2 in the growth environments quickly and expresses the O_2 responsive genes. In this study, we investigated the function of O_2-responsive nror operon enzymes by purification of the proteins and characterization of their enzymatic activity. We propose that the obligate anaerobe C. acetobutylicum possesses an efficient multi-enzyme complex that can scavenge O_2 and ROS using NROR as a master electron donor protein. The primary factor responsible for aerobic growth inhibition of Bifidobacterium species is proposed to be the production of H_2O_2 in the growth medium. A H_2O_2-forming NADH oxidase was purified from O_2-sensitive Bifidobacterium bifidum and was identified as a b-type DHOD. The kinetic parameters suggested that the enzyme could be involved in H_2O_2 production in highly aerated environments.

我们的目的是了解必需厌氧菌梭状芽孢杆菌和双歧杆菌对氧气敏感的机制。乙酰丁酸梭菌是一种高效的丙酮丁醇发酵菌，能快速感知生长环境中的O2污染，并表达O2应答基因。在这项研究中，我们通过纯化蛋白质和鉴定它们的酶活性来研究氧响应的NOR操纵子酶的功能。我们认为专性厌氧菌乙酰丁酸杆菌具有高效的多酶复合体，可以利用NROR作为主要的电子供体蛋白来清除O2和ROS。双歧杆菌好氧生长抑制的主要因素是生长介质中H_2O_2的产生。从双歧双歧杆菌中分离纯化出一种产H_2O_2的NADH氧化酶，经鉴定为b型DHOD。动力学参数表明，该酶可能参与了高曝气环境中过氧化氢的产生。

项目成果

Crystallization and Preliminary X-ray Analysis of NADH:rubredoxin Oxidoreductase from Clostridium acetobutylicum

丙酮丁醇梭菌 NADH:红氧还蛋白氧化还原酶的结晶和初步 X 射线分析

• DOI: 10.1107/s1744309109047162
• 发表时间: 2010
• 期刊: Acta Crystallogr
• 作者: K. Nishikawa;Y. Shomura;S. Kawasaki;Y. Niimura and Y. Higuchi
• 通讯作者: Y. Niimura and Y. Higuchi

乳酸菌培養菌体による過酸化物分解(第9報)-菌体外過酸化水素を分解するp. pentosaceus Bel株の過酸化物分解酵素-

培养乳酸菌细胞的过氧化物分解（第9次报告） - 分解细胞外过氧化氢的戊糖丙酸杆菌Bel菌株的过氧化物降解酶 -

• 发表时间: 2009
• 作者: 澤井仁美;杉本宏;加藤康夫;浅野泰久;城宣嗣;青野重利;永尾寿浩;濱田 恭宏
• 通讯作者: 濱田 恭宏

Mechanism of growth inhibition by oxygen in Bifidobacterium species

氧气抑制双歧杆菌生长的机制

• 发表时间: 2010
• 作者: 加藤康夫;大塚耕太郎;松村航;荻田信二郎;川崎信治
• 通讯作者: 川崎信治

嫌気性細菌の酸素適応機構

厌氧菌的氧适应机制

• 发表时间: 2009
• 作者: 荻田信二郎;野村泰治;加藤康夫;金子千晶;永尾寿浩;加藤康夫;川崎信治
• 通讯作者: 川崎信治

O2 and Reactive Oxygen Species Detoxification Complex, Composed of O2-Responsive NADH:Rubredoxin Oxidoreductase-Flavoprotein A2-Desulfoferrodoxin Operon Enzymes, Rubperoxin, and Rubredoxin, in Clostridium acetobutylicum

• DOI: 10.1128/aem.01425-08
• 发表时间: 2009-02-15
• 期刊: APPLIED AND ENVIRONMENTAL MICROBIOLOGY
• 影响因子: 4.4
• 作者: Kawasaki, Shinji;Sakai, Yu;Niimura, Youichi
• 通讯作者: Niimura, Youichi