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右旋糖酐对Savinase蛋白酶的化学修饰及酶学性质研究
引用本文:朱怡然,范雪荣,王强,陈忍忍,余圆圆,袁久刚.右旋糖酐对Savinase蛋白酶的化学修饰及酶学性质研究[J].中国生物工程杂志,2011,31(10):45-49.
作者姓名:朱怡然  范雪荣  王强  陈忍忍  余圆圆  袁久刚
作者单位:江南大学生态纺织教育部重点实验室 无锡 214122
基金项目:国家"863"计划,国家自然科学基金,江苏省高等学校优秀科技创新团队资助项目
摘    要:采用经高碘酸钠活化的右旋糖酐修饰Savinase蛋白酶,通过凝胶过滤层析(GPC)和圆二色性光谱(CD)表征了修饰后蛋白酶分子量和结构的变化,测试了修饰酶的反应动力学参数,并考察了温度及pH对修饰酶活力的影响。凝胶过滤层析结果证明修饰后蛋白酶分子量明显提高,圆二色光谱分析表明修饰后蛋白酶的结构有所改变,进一步验证了右旋糖酐和蛋白酶发生了反应。与原酶相比,修饰酶对底物的亲和力增加。原酶和修饰酶的最适温度均为40℃,在30℃~50℃之间修饰酶表现出优于原酶的热稳定性。在pH8.5~9.5之间,修饰酶的稳定性高于原酶。

关 键 词:Savinase蛋白酶  右旋糖酐  化学修饰  分子量  
收稿时间:2011-04-29
修稿时间:2011-06-03

Chemical Modification of Savinase by Dextran and Its Enzyme Properties
ZHU Yi-ran,FAN Xue-rong,WANG Qiang,CHEN Ren-ren,YU Yuan-yuan,YUAN Jiu-gang.Chemical Modification of Savinase by Dextran and Its Enzyme Properties[J].China Biotechnology,2011,31(10):45-49.
Authors:ZHU Yi-ran  FAN Xue-rong  WANG Qiang  CHEN Ren-ren  YU Yuan-yuan  YUAN Jiu-gang
Institution:(Jiangnan University Key Laboratory of Eco-Textiles,Ministry of Education,Wuxi 214122,China)
Abstract:Savinase was chemically modified using NaIO4 oxidised dextran. The molecular size and the secondary structure of modified Savinase were characterized by GPC and CD. The factors related with the activity of the modified Savinase, such as kinetic constant, temperature and pH value were studied and compared with those of native Savinase. GPC results showed that the molecular size of the protease increased after chemical modification. CD spectra revealed the difference between native and modified Savinase, which further demonstrated the conjugate of oxidized dextran and Savinase. Compared with the native Savinase, the modified one had a higher affinity to casein. The optimum reaction temperature for both of the protease was 40℃, and the modified Savinase had a better thermal stability at 30℃~50℃. The stability of modified Savinase is better than the native one at pH8.5~9.5.
Keywords:Savinase  Dextran  Chemical modification  Molecular size
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