食品科学 ›› 2010, Vol. 31 ›› Issue (1): 181-184.doi: 10.7506/spkx1002-6630-201001042

• 生物工程 • 上一篇    下一篇

鳀鱼胰蛋白酶分离纯化及性质初步研究

吕英涛1,康从民1,*,王 琳1,薛长湖2   

  1. 1.青岛科技大学化工学院 2.中国海洋大学食品科学与工程学院
  • 收稿日期:2009-02-16 修回日期:2009-06-16 出版日期:2010-01-01 发布日期:2014-05-19
  • 通讯作者: 康从民 E-mail:cmkang@qust.edu.cn
  • 基金资助:

    山东省自然科学基金项目(Y2007C176;Y2007C175);山东省教育厅科技计划资助项目(J07YC08);
    青岛市科技计划基础研究资助项目(09-1-3-74-JCH);山东省高等学校科技计划项目(J09LC20)

Separation, Purification and Characterization of Trypsin from Anchovy

LU Ying-tao1,KANG Cong-min1,*,WANG Lin1,XUE Chang-hu2   

  1. (1. College of Chemical Engineering, Qingdao University of Science and Technology, Qingdao 266042, China;
    2. College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China)
  • Received:2009-02-16 Revised:2009-06-16 Online:2010-01-01 Published:2014-05-19
  • Contact: KANG Cong-min1,*, E-mail:cmkang@qust.edu.cn

摘要:

鳀鱼内源蛋白酶粗酶中主要含有4 种蛋白酶,其中活性最强的为胰蛋白酶,其对鳀鱼自溶所起的作用最大。鳀鱼蛋白酶粗酶经过两次Q-Sepharose FF 离子交换色谱和一次Sephacryl S-300 凝胶色谱分离,得到纯化的鳀鱼胰蛋白酶,对分离纯化后鳀鱼胰蛋白酶生化特性进行初步研究。结果表明,最适pH9.0,最适温度为55℃,并且有非常好的热稳定性。鳀鱼胰蛋白酶能分解胰蛋白酶的特异性底物,也可以分解天然蛋白,钙、镁离子都对胰蛋白酶酶活有一定的稳定作用。

关键词: 鳀鱼自溶酶, 胰蛋白酶, 分离纯化, 特性

Abstract:

Anchovy endogenous crude protease is mainly composed of four proteases. Trypsin plays the most important role in reactions with casein. Trypsin-like protease was purified using a combined separation method of ion exchange chromatography and gel filtration chromatography. Preliminary characterization of purified trypsin was also conducted. Results suggested that the purified trypsin from anchovy exhibited excellent thermal and pH stability. The optimal pH and temperature of this enzyme was 9.0 and 55 ℃, respectively. It also exhibited high substrate specificity. According to inhibition profile of specific protease inhibitors and metal ions, trypsin was a kind of metal-activated enzyme rather than a kind of metalloenzyme.

Key words: anchovy endogenous proteases, trypsin, separation and purification, characterization

中图分类号: