食品科学 ›› 2005, Vol. 26 ›› Issue (2): 59-63.

• 基础研究 • 上一篇    下一篇

纳豆激酶分离纯化及纤溶活性研究

 王萍, 陈钧, 杨小明, 鲍艳霞   

  1. 江苏大学生环学院制药工程系; 江西农业大学动物科学技术学院;
  • 出版日期:2005-02-15 发布日期:2011-09-19

Research on Purification and Fibrinolytic Activity Assay of Nattokinase from Natto

 WANG  Ping, CHEN  Jun, YANG  Xiao-Ming, BAO  Yan-Xia   

  1. 1.School of Biological and Enviromental Engineering,Jiangsu University; 2.College of Animal Science and Technology,Jiangxi Agriculture University
  • Online:2005-02-15 Published:2011-09-19

摘要: 经过生理盐水浸提、(NH4)2SO4分级沉淀、Sephadex G-100凝胶层析等纯化步骤,从纳豆中获得层析纯的纳豆激酶,纯化倍数15.6,回收率10.2%,SDS-PAGE电泳显示为二个组分,分子量在29000左右。对其纤溶性质研究表明:纳豆激酶活性的最适pH为8.0,pH6~10溶液中40℃以下基本稳定,pH<5失去纤溶活性;模拟胃环境的酸性条件下,粗酶失去纤溶活性,而纳豆生理盐水浸提液纤溶活性保持25%;SDS-PAGE电泳显示,胰蛋白酶对纳豆激酶成分没有降解作用;体外溶栓作用表明,纳豆激酶溶解纤维蛋白的方式是直接溶解,而不是通过激活纤溶酶原。

关键词: 纳豆激酶, 分离纯化, 纤溶活性

Abstract: The nattokinase was purified from natto through procedures of saline extraction, ammonium sulfate precipitation, and SephadexG-100 gel chromatograph purification with a fold of 15.6 and overall yield of 10.2%. The purified enzyme contained two protein components with molecular weight about 29kD by SDS-PAGE. Properties of fibrinolytic activity showed nattokinase. Its optimum pH was 8, stable within pH 6~10 and below 40℃, but inactivated completely below pH5. Under simulating acid environment of stomach, the fibrinolytic activity of crude extract of nattokinase lost completely while 25% of the saline extract of natto retained. The SDS-PAGE results showed that the nattokinase could not be hydrolyzed by trypsin. Nattokinase could hydrolyze fibrin directly rather than by activating plasminogen in vitro.

Key words: nattokinase, purification, fibrinolytic activity