J4

• 生命科学 • 上一篇    下一篇

以卵清蛋白为载体的固定化木瓜蛋白酶的制备及其性质

黄宜兵, 吴晓霞, 吴胜男, 徐力, 张学忠   

  1. 吉林大学分子酶学工程教育部重点实验室, 长春 130023
  • 收稿日期:2004-02-27 修回日期:1900-01-01 出版日期:2004-10-26 发布日期:2004-10-26
  • 通讯作者: 张学忠

Preparation and its properties of the immobilized papain with\= denatured egg white protein as the support

HUANG Yi-bing, WU Xiao-xia, WU Sheng-nan, XU Li, ZHANG Xue-zhong   

  1. Key Laboratory of Molecular Enzymology and Engineering, Ministry of Education, Jilin University, Changchun 130023, China
  • Received:2004-02-27 Revised:1900-01-01 Online:2004-10-26 Published:2004-10-26
  • Contact: ZHANG Xue-zhong

摘要: 以变性的卵清蛋白为载体, 用戊二醛为交联剂固定木瓜蛋白酶. 通过正交实验考察了固定化pH值、 交联剂戊二醛浓度、 交联时间和酶用量对固定化木瓜蛋白酶活力的影响,确定了最适固定化条件: 5%戊二醛, pH 6.0, 交联时间20 h, 每克载体含酶量12 mg. 对比研究了游离木瓜蛋白酶和固定化木瓜蛋白酶的性质, 所得到的固定化木瓜蛋白酶最适温度90 ℃, 最适pH 9.0, 米氏常数53.6 mg/mL, 其热稳定性及耐热性均较游离酶显著提高.

关键词: 木瓜蛋白酶, 固定化, 卵清蛋白, 戊二醛

Abstract: Papain was immobilized on a denatured egg white protein as the support with glutaraldehyde as the cross-linker. The effects of pH, the concentration of glutaraldehyde, time and the amount of the enzyme on the activity of the immobilized papain were examined through the experiments of orthogonal test. The optimal condition for the immobilization was set up, i.e., glutaraldehyde concentration is 5%; pH 6.0; the quantity of enzyme used 12 mg/(g support) and the reaction time 20 h. At the same time, the comparative study on the properties of free and immobilized p apain was made. The optimal temperature and pH are 90 ℃ and 9.0, respectively . The Km for the immobilized enzyme is 53.6 mg/mL. The thermostability and thermotolerance of the immobilized enzyme are obviously increased compared to those of the free one.

Key words: papain, immobilization, egg white protein, glutaraldehyde

中图分类号: 

  • Q55