DIOL DEHYDRATASE AND DIOL METABOLISM IN CLOSTRIDIUM GLYCOLICUM

乙二醇梭菌中的二醇脱水酶和二醇代谢

• 批准号: 3966514
• 负责人: M G HARTMANIS
• 金额: --
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3966514
• 关键词: Clostridium; cofactor; diol; ethylene glycols; fermentation; hydro lyase; microorganism metabolism; propylene glycols; proteolysis

The oxygen sensitive and membrane bound diol dehydratase from Clostridium glycolicum was solubilized from its matrix by sonication of crude membrane preparations anaerobically in 0.1 M CHES buffer, pH 8.5 or 9.0, containing 2 mM dithiothreitol. Treatment with organic solvents, a variety of ionic and nonionic detergents, high ionic strength, or phospholipase A did not solubilize any diol dehydratase activity. Addition of 30% dimethylsulfoxide and 0.15 mg/ml of lysophosphatidylcholine to the CHES buffer before sonication markedly increased recovery of enzyme activity. Up to 45% of the activity could be recovered after centrifugation for 1 h at 105,000 x g. This solubilization method was also shown to work for the membrane bound formate dehydrogenase from E. coli. More than 95% of the activity was recovered in the supernatant after sonication and centrifugation.

梭状芽胞杆菌氧敏膜结合二醇脱水酶 粗膜超声可使乙交酯从其基质中溶解 在0.1M CHES缓冲液中厌氧制剂，pH 8.5或9.0，含有 2 mM二硫苏糖醇。用有机溶剂处理，各种离子 而非离子洗涤剂、高离子强度或磷脂酶A则没有 溶解任何二醇脱水酶的活性。增加30% 二甲基亚砜和0.15 mg/ml溶血磷脂酰胆碱对CHS的作用 超声前缓冲液显著提高了酶活力的恢复。 离心1h后，酶活可恢复到45% 在105,000×g。这种增溶方法也被证明对 大肠杆菌膜结合型甲酸脱氢酶。超过95%的人 超声处理后上清液中活性恢复， 离心法。