用醇类从酵母菌中释放超氧化物歧化酶

研究了一种用醇类有机溶剂处理法从酵母菌中选择性地释放超氧化物歧化酶(SOD)的方法。当采用异丙醇浓度为90％,浸泡120min,抽提缓冲液为50mmol/L磷酸盐缓冲液(Ph7.0)时,抽提20h,SOD释放的活力为300u/ml,杂蛋白释放量最少,SOD比活达300u/mg。SOD释放率可达90％,和传统的超声波法和机械法相比,而比活提高了25倍。这种方法不需要任何复要设备,操作简单,成本低廉,在释放SOD同时,可达到初步纯化SOD的效果。

Selective Release of Superoxide Dismutase from Saccharomyces cerevisiae by Alcoholic Solvent

A new and cheap approach in which the alcoholic solvents were used to release selectively superoxide dismutase (SOD) from Saccharomyces cerevisiae, was developed. The effects, which influenced the release of SOD, such as the alcoholic slovent type, concentration, treating time and the extractive buffer concentraton, extractive time, pH were studied. When 90% isopropanol was used to treat the cells for 120 min and then 50mmol/L phosphate buffer (pH7. 0) was added after removal of solvent by filtration, the SOD released was up to 300u/ml with specific activity 300u/mg. The recovery of SOD is 90%, comparing to traditional methods such as ultrasonic approach, while the specific activity of SOD increasd 25 folds. The new approach does not require any complicated device and is a simple mild and inexpeneive method of intracellular enzyme release, where the target enzyme SOD can by purified simultaneously.