贵阳腐霉类枯草杆菌蛋白酶(Pr1)的纯化与部分特性研究

目的:纯化灭蚊真菌贵阳腐霉胞外蛋白酶类枯草杆菌蛋白酶,并测定其分子量.方法:培养贵阳腐霉菌丝体,用几丁质诱导,使其产生大量胞外酶,对其产酶条件进行了部分优化.将粗酶液经过脱盐,浓缩进一步纯化,再过Sephadex G-100柱子层析并进行SDS-PAGE,分离类枯草杆菌蛋白酶.结果:(1)以几丁质为唯一碳氮源,在26℃,初始pH值6.0,1%的菌丝接种量为最佳诱导条件;(2)分离得到的类枯草杆菌蛋白酶,并测定其分子量约为33kDa.结论:类枯草杆菌蛋白酶纯化成功,结果较理想,为进一步研究提供依据.

Purification and Characterization of A Subtilisin-like Protease (Pr1) of Pythium guiyangense

Objective: To induce and purify a subtilisin-like protease ( Pr1 ) of Pythium guiyangense, a mosquito-killing fungus, and to learn the molecular weight of the Pr1. Methods: Hyphae of P. guiyangense were cultured in liquid KPYG2 medium for 5 days before being shaked with chitin for 30 hours. The suspension was, then, collected, filtered and concentrated. Pr1 was isolated and purified from the suspension by passing it through Sephadex G-100 column, and running SDS-PAGE. Results: (1) The optimum conditions for protease induction were: 26 ℃, initial pH 6.0 and inoculative quantity of fungal hyphae = 1 g/100 ml, using chitin as the inducer; (2) Pr1 was purified and its molecular weight was determined as 33kDa. Conclusion: The induction, purification and molecular weight determination of Pr1 are successful and the results provide useful basis for further study.