纳豆激酶的分离纯化及酶学性质研究  

Study on purification and enzyme kinetics of nattokinase

作　　者：董志奎[1] 杨超[1] 尹宗宁[1] 李萌[1] 

机构地区：[1]四川大学华西药学院,四川成都610041

出　　处：《中国生化药物杂志》2009年第5期298-301,共4页Chinese Journal of Biochemical Pharmaceutics

基　　金：国家自然科学基金项目(30371696)

摘　　要：目的研究纳豆激酶分离纯化工艺及酶学性质。方法纳豆激酶发酵液的粗提物经Superdex 75凝胶色谱和聚丙烯酰胺凝胶电泳(PAGE)分离纯化,采用TAME法测定酶的活性,通过SDS-PAGE对纯化结果进行了检验。结果SDS-PAGE中显示单一色带,相对分子质量28000,以TAME为底物时纳豆激酶的米氏常数(Km)为35.47mmol/L,最适宜的温度37℃,最适宜pH为8.6。结论该分离纯化方法可以得到较纯的纳豆激酶。Purpose To study the techniques of the separation and purification of the nattokinase and its characterization. Methods The natuokinase extract was purified by gel-chromatography（Superdex75） and polyacrylamide gel electrophoresis（PAGE）. The purified nattokinase was tested by SDS-PAGE, and the TAME method was used to determine the activity of the enzyme. Results There was only one strip in the SDS-PAGE atlas, and the molecular weight of nattokinase was 28 kDa. The Miehaelis constant（Km） of nattokinase was 35.47 mmol/L when TAME was the substrate, and the best reaction temperature was 37℃ and the best reaction pH was 8.0. Conclusion Purified nattokinase can be obtained by the the techniques of the separation and purification.

关 键 词：纳豆激酶 分离纯化 酶学性质 米氏常数(Km) 

分 类 号：Q55[生物学—生物化学] TQ464.8[化学工程—制药化工]