食品科学

• 生物工程 • 上一篇    下一篇

白姑鱼胰蛋白酶的分离纯化及其性质分析

李 欢,李 婷,詹云超,杜翠红*   

  1. 集美大学食品与生物工程学院,水产品深加工技术国家地方联合工程研究中心,福建 厦门 361021
  • 出版日期:2016-07-15 发布日期:2016-07-26
  • 通讯作者: 杜翠红
  • 基金资助:

    厦门市海洋渔业局(南方海洋中心)项目(14CZP03HJ04);福建省科技厅引导性项目(2016N0020)

Purification and Characterization of Trypsin from White Croaker (Argyrosomus argentatus)

LI Huan, LI Ting, ZHAN Yunchao, DU Cuihong   

  1. National & Local Joint Engineering Research Center of Processing Techonology for Aquatic Products, College of Food and Biological
    Engineering, Jimei University, Xiamen 361021, China
  • Online:2016-07-15 Published:2016-07-26
  • Contact: DU Cuihong

摘要:

本研究从白姑鱼幽门垂组织中提取胰蛋白酶粗酶,然后经硫酸铵分级沉淀、DEAE-Sepharose Fast Flow阴离子交换层析及Sephacryl S-200凝胶过滤层析等分离纯化技术,得到一种阴离子型胰蛋白酶(命名为:Trypsin A),并对该阴离子型胰蛋白酶进行鉴定及性质分析。结果表明:Trypsin A的分子质量约为28 kD,其最适反应温度为50 ℃,能在低于65 ℃条件下保持稳定;最适pH值为9.5,酸碱耐受范围为pH 9.5~11.0;Western blotting分析表明,白姑鱼阴离子型胰蛋白酶可以与抗鲫鱼胰蛋白酶抗体反应,符合胰蛋白酶活性区域的高度保守性;丝氨酸类蛋白酶抑制剂对白姑鱼胰蛋白酶具有抑制作用。

关键词: 白姑鱼, 胰蛋白酶, 分离纯化, 酶学性质

Abstract:

An anionic trypsin (named as trypsin A) from the pyloric caeca of white croaker (Argyrosomus argentatus) was
purified by ammonium precipitation, DEAE-Sepharose Fast Flow anion exchange chromatography and Sephacryl S-200 gel
filtration chromatography. The characteristics of trypsin A were investigated. The results showed that trypsin A migrated as
a single protein band with a molecular weight of about 28 kD in SDS-PAGE. Trypsin A exhibited its optimal temperature at
50 ℃ and was stable below 65 ℃. Trypsin A revealed its optimal pH at 9.5 and was stable in the pH range from 9.5 to 11.0.
The purified trypsin A was analyzed by western blotting using anti-common carp trypsin antibody, and the result showed
that the active region of the trypsin was highly conservative. Serine proteinase inhibitors were effective against trypsin A.

Key words: white croaker, trypsin, separation and purification, enzymatic characteristics

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