哈茨木霉S30胞外几丁质酶的纯化及特性  

Purification and Properties of an Extracellular Chitinase Produced by Trichoderma harzianum S30

作　　者：张世宏[1] 李多川[1] 刘开启[1] 魏毅 

机构地区：[1]山东农业大学植保系,泰安271018 [2]山东省棉花研究中心,济南250100

出　　处：《中国生物防治》2000年第3期107-110,共4页Chinese Journal of Biological Control

基　　金：国家自然科学基金!(3970 0 0 97);国际科学基金!(C/ 2 80 1- 1)资助项目

摘　　要：哈茨木霉 S30在 SMCS液体培养基中恒温摇床 ( 2 0 0 r/ min,2 7.5℃ )上培养 1 5天 ,培养滤液经硫酸铵分级沉淀、DEAE- Sepharose Fast Flow阴离子交换柱层析、Phenyl- SepharoseFast Flow疏水层析、CM- Sepharose Fast Flow阳离子柱层析、Phenyl- Sepharose HP疏水柱层析获得了凝胶电泳谱带单一的几丁质酶。几丁质酶反应的最适温度为 40℃ ,最适 p H值为5 .0。 70℃下保温 1 0分钟后 ,该酶仍保持一定剩余活力 ,说明其耐温性较强。金属离子存在对酶活性有显著影响 ,Mg2 +、Ba2 +、Mn2 +对酶有激活作用 ,其中以 Mg2 +作用最强。An extracellular chitinase was purified to SDS PAGE by ammonium sulfate fraction, DEAE Sepharose Fast Flow chromatography, Phenyl Sepharose Fast Flow chromatography, CM Sepharose Fast Flow chromatography and Phenyl Sepharose High Performance (1×1ml) chromatography when Trichoderma harzianum S30 was grown in SMCS broth at 27 5℃ on a rotary shaker at 200r/min for 15 d The optimum pH for the enzyme was pH 5 0, and maximum activity was obtained at the temperature of 40℃ The chitinase was thermostable at 40℃ The enzyme activity was reduced to 69 32%, 33 07%, 8 33% activity after 10 min at 50℃, 60℃ and 70℃, respectively The addition of Mg 2+ , Mn 2+ or Ba 2+ increased the enzyme activity, and Mg 2+ was the bestn

关 键 词：哈茨木霉S30 几丁质酶 纯化 特性 

分 类 号：Q557.02[生物学—生物化学]