摘要
有氧条件下,建立了克雷伯杆菌破碎方法及其生产1,3-丙二醇代谢途径中关键酶酶活测定方法。超声波细胞破碎优化条件为:超声频率30kHz,100%振幅,间歇频率0.6,超声时间12min。细胞抽提液中甘油脱氢酶、1,3-丙二醇氧化还原酶、甘油脱水酶酶活测定的最适pH分别为12、9.5和7.0,最适温度分别为45oC、45oC和37 。甘油脱氢酶、1,3-丙二醇氧化还原酶用初速度法测定。甘油脱水酶用终止法测定。
Under aerobic conditions the determination method of key enzyme activity of 1,3-propanediol production by Klebsiella pneumoniae was established. The optimal conditions of cell disruption by ultrasonic wave were frequency of 30kHz, amplitude of 100%, intermittent frequency of 0.6 and ultrasonic time of 12min. The optimal pH of determination of the activities of glycerol dehydrogenase, 1,3- propanediol oxidoreductase and glycerol dehydratase in cell-free extract were 12, 9.5 and 7.0 and the optimal temperature of those were 45oC , 45oC and 37℃. The activities of glycerol dehydrogenase and 1,3-propanediol oxidoreductase were assayed by the method of initial velocity and the activity of glycerol dehydratase was determined by endpoint measurement.
基金
国家自然科学基金(20276026)
福建省自然科学重点项目基金资助(D0120002)
