Molecular breeding of a toxic chemicals-degrading-bacterium

有毒化学品降解菌的分子育种

• 批准号: 05660112
• 负责人: UCHIYAMA Hiroo
• 金额: $ 1.34万
• 依托单位: National Institute for Environmental Studies
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05660112/
• 关键词: methane monooxygenase; methanotroph; trichloroethylene

1. Soluble methane monooxygenase (sMMO) gene was cloned from a methane-utilizing bacterium, Methylocystis sp.M (strain M), and sequenced.(1) sMMO structural gene (about 6kb) and its promotor region were sequenced, and those were confirmed to be sMMO genes by comparing them with complemental DNA sequences derived from N-terminal amino acid sequences of purified sMMO in strain M.(2) Some characteristics were cleared as follows ;・ sMMO from strain M was related closely with those of methylosinus trichosporium OB3b and Methylococcus capsulatus Bath in DNA sequences and putative amino acid sequences.・ Metal-binding site in a active center of sMMO,high G/C content in the third letter of the used codons, and prothsetic groups binding site were confirmed in that DNA sequence.2. Plasmid pWM3 coding sMMO genes was introduced into E.coli. Trichloroethylene degradation activity was not detected in E.coli harboring pWM3, and no difference in lysates was observed between E.coli harboring pWM3 and host by SDS-PAGE.Hydroxylase, one of the components of sMMO,consisted with alpha 2 beta 2 gamma 2 subunits and requires ferric ion in the active center, indicating that the precise assembly of those subunits and a metal would be required to express hydroxylase activity.

1。从甲烷利润细菌，甲基固醇sp.M（M）和测序的甲烷脱氧基因（SMMO）基因中克隆。（1）SMMO结构基因（约6KB）（约6KB）及其启动子区域及其序列被确认为SMMO GENES的序列，并被序列序列序列序列序列序列序列，并被序列序列序列序列序列序列序列序列，并被序列序列序列序列的NNA序列序列序列，DNA序列是DNA序列的dNA序列。菌株M中的SMMO（2）（2）如下清除了一些特征；从M菌株中的SMMO与DNA序列中的DNA序列中的Trichosinus porporium ob3b和甲基甲虫囊浴的甲基甲基孢子菌和甲基甲状腺胶菌的相关密切相关。在该DNA序列中2。将质粒PWM3编码SMMO基因引入了大肠杆菌。 Trichloroethylene degradation activity was not detected in E.coli harboring pWM3, and no difference in lysates was observed between E.coli harboring pWM3 and host by SDS-PAGE.Hydroxylase, one of the components of sMMO,consisted with alpha 2 beta 2 gamma 2 subunits and requires ferric ion in the active center, indicating that the precise assembly of those subunits and a将金属表达羟化酶活性。

项目成果

内山裕夫: "微生物機能の多様性" 別府輝彦, 526 (1995)

内山博夫：“微生物功能的多样性” Teruhiko Beppu，526（1995）

内山裕夫: "微生物機能の多様性-メタン資化性菌と環境浄化-" 別府輝彦 編 学会出版センター, 526 (1995)

内山博夫：《微生物功能的多样性——甲烷同化细菌与环境净化》，别府照彦编，学会出版中心，526（1995）

内山裕夫・矢木修身: "揮発性有機塩素化合物を分解する微生物" バイオサイエンスとインダストリー. 52. 879-884 (1994)

Hiroo Uchiyama 和 Osumi Yagi：“分解挥发性有机氯化合物的微生物”生物科学与工业 52. 879-884 (1994)。

H.Uchiyama: Methane-utilizing bacteria and remediation of polluted site In : Biodiversity of Bacterial Functions T.Beppu (ed). Gakkai-Shuppan-Center, 43-56 (1995)

H.Uchiyama：利用甲烷的细菌和污染场地的修复，见：细菌功能的生物多样性 T.Beppu（编辑）。

内山裕夫: "揮発性有機塩素化合物を分解する微生物" バイオサイエンスとインダストリー. 52. 879-884 (1994)

Hiroo Uchiyama：“分解挥发性有机氯化合物的微生物”生物科学与工业 52. 879-884 (1994)。