Molecular Understanding of the Coryne-form Bacteria and its Industrial Application

棒状细菌的分子认识及其工业应用

基本信息

批准号：
11356003
负责人：
TOMTA Fusao
金额：
$ 24.26万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

项目摘要

(1) TOMITA: H^+-ATPase operon genes of Corynebacterium glutamicum were cloned and sequenced. The transformant carrying the atp operon genes showed 2.5-fold higher ATPase activities than the wild type. The transformant did now exhibit a distinct difference in fermentation characteristics. However, the maximum levels of both growth and glutamic acid production were somewhat higher than those of the wild type. Also in the glutamic acid production phase, the transformant showed higher glucose consumption rate, respiration rate, membrane potential, internal pH value than those of the wild type.(2) MATSUSHITA: Membrane vesicles were successfully prepared from Corynebacterium glutamicum, and energetics of the respiratory chain was analyzed by using the membrane vesicles thus prepared. Furthermore, NADPH oxidase, characteristic in the respiratory chain, was purified and shown to be an NADH dehydrogenase II (ndh), of which the function in the respiratory chain and energy metabolism was examined … More by constructing ndh-disrupted strain.(3) KATSUMATA: Alanine dehydrogenase gene of Corynebacterium glutamicum was cloned and sequenced. A transformant of C. glutamicum expressing the gene was obtained. However, in C. glutamicum, alanine biosynthesis appeared to be performed by transaminase. The amplification of the transaminase gene resulted in the production of alanine 4.5 times higher than that of the parent.(4) NAGAI: Peptidoglycan synthesis genes, murI, murC, murE, and ftsI, were isolated form coryneform bacteria. ltsA gene encoding a novel amido-transferase involved in the formation of cell surface structure was isolated from Corynebacterium glutamicum by the analysis of lysozyme sensitive mutant. Defects of the ltsA gene induced production of L-glutamic acid.(5) NAKAMORI: Cysteine metabolism of Brevibacterium flavum (Corynebacterium glutamicum) was investigated and cysteine desulfuhydrase (CD) of this strain was identified. Cysteine production with CD disruptant was performed. Serine acetyltransferase gene of this strain was also identified and characterized.(6) KUMAGAI: Cloning of γ -glutamyltranspeptidase (GGT) gene of Corynebacterium was carried out but not successful. Active site of E. coli GGT was determined and autoprocessing mechanism was elucidated, α -Galactosidase of Bifidobacterium, a kind of Corynebacterium, was characterized and the gene was cloned. Less

(1)TOMITA：对谷氨酸棒杆菌的H^+-ATP酶操纵子基因进行了克隆和测序，携带atp操纵子基因的转化体显示出比野生型高2.5倍的ATP酶活性。现在，转化体在发酵特性方面表现出明显的差异。然而，在谷氨酸生产阶段，转化体的生长和谷氨酸生产的最高水平也稍高于野生型。表现出比野生型更高的葡萄糖消耗率、呼吸率、膜电位、内部pH值。(2)松下：从谷氨酸棒杆菌中成功制备膜囊泡，并利用膜囊泡分析呼吸链的能量学此外，呼吸链中的特征性 NADPH 氧化酶被纯化并显示为 NADH 脱氢酶 II (ndh)，其在呼吸链中的功能(3) KATSUMATA：克隆并测序了谷氨酸棒杆菌的丙氨酸脱氢酶基因，并获得了表达该基因的谷氨酸棒杆菌转化体。丙氨酸生物合成似乎是由转氨酶进行的。转氨酶基因的扩增导致丙氨酸的产生。比亲本高4.5倍。(4)NAGAI：从棒状细菌中分离出肽聚糖合成基因murI、murC、murE和ftsI，ltsA基因编码参与细胞表面结构形成的新型酰胺转移酶。通过分析溶菌酶敏感突变体从谷氨酸棒杆菌中分离出 ltsA 基因的缺陷诱导的生产。 (5) NAKAMORI：研究了黄色短杆菌（谷氨酸棒杆菌）的半胱氨酸代谢，并鉴定了该菌株的半胱氨酸脱硫酶（CD）与 CD 丝氨酸乙酰转移酶基因的生产。 (6)KUMAGAI: γ-谷氨酰转肽酶(GGT)基因的克隆进行了棒状杆菌的鉴定，但没有成功，确定了大肠杆菌GGT的活性位点并阐明了自加工机制，对棒状杆菌双歧杆菌的α-半乳糖苷酶进行了表征并克隆了该基因。