于声,段斯亮,苏晓庆
YU Sheng,DUAN Siliang,SU Xiaoqing(Department of Biology

• 1:贵阳医学院生物学教研室
• 2:贵阳医学院生物学教研室
• 3:贵阳医学院生物学教研室 贵州贵阳550004
• 4:贵州贵阳550004

摘要(Abstract)：

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。
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.

关键词(KeyWords)： 贵阳腐霉;类枯草杆菌蛋白酶;纯化;分子量;蚊虫生物防治
Pythium guiyangense;subtilisin-like protease;purification;molecular weight;mosquito biocontrol

Abstract:

Keywords:

基金项目(Foundation): 贵州省优秀科技教育人才省长资金;; 贵州省高层次人才科研条件特助经费资助

作者(Author): 于声,段斯亮,苏晓庆
YU Sheng,DUAN Siliang,SU Xiaoqing(Department of Biology

DOI: 10.19367/j.cnki.1000-2707.2007.02.006

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