食品科学 ›› 2009, Vol. 30 ›› Issue (1): 186-189.doi: 10.7506/spkx1002-6630-200901044

• 生物工程 • 上一篇    下一篇

定向固定化葡萄糖氧化酶及其酶学性质的研究

周建芹1,陈韶华2,王剑文1   

  1. 1.苏州大学医学部药学院 2.苏州大学医学部实验中心
  • 收稿日期:2007-12-22 修回日期:2008-05-08 出版日期:2009-01-01 发布日期:2010-12-29
  • 通讯作者: 周建芹 E-mail:zkzhu@ustc.edu
  • 基金资助:

    江苏省自然科学基金项目(BK2007051)

Study on Oriented Immobilization of Glucose Oxidase and Its Enzymatic Properties

ZHOU Jian-qin1,CHEN Shao-hua2,WANG Jian-wen1   

  1. (1. School of Pharmacy, Medical College, Soochow University, Suzhou 215123, Chin;
    2.Experiment Center of Medical College, Soochow University, Suzhou 215123, China)
  • Received:2007-12-22 Revised:2008-05-08 Online:2009-01-01 Published:2010-12-29
  • Contact: ZHOU Jian-qin1 E-mail:zkzhu@ustc.edu

摘要:

戊二醛将伴刀豆球蛋白(ConA)和载体壳聚糖膜交联,然后利用ConA 与葡萄糖氧化酶糖链的特异性结合作用,实现酶的定向固定化。定向固定化的最适条件为戊二醛浓度0.1%、ConA 浓度0.02mg/ml、葡萄糖氧化酶浓度0.08mg/ml。定向固定化葡萄糖氧化酶的最适pH4.0、最适温度57℃,米氏常数Km 为15.84mmol/L,与游离酶及非定向固定化葡萄糖氧化酶比较,定向固定化葡萄糖氧化酶的最适pH 值向酸性范围发生了偏移并有更宽的pH 值适用范围,最适温度提高,与底物的亲和力较大。

关键词: 葡萄糖氧化酶, 伴刀豆球蛋白, 定向固定化

Abstract:

Concanavalin A (ConA), which has a very strong interaction with glycoprotein, was immobilized on preactivated glutaraldehyde-modified chitosan microspheres, and then oriented immobilization of glucose oxidase was carried out on the strong interaction. The optimal immobilization conditions are as follows: glutaraldehyde concentration 0.1%, ConA concentration 0.02 mg/ml and glucose oxidase concentration 0.08 mg/ml. The optimum pH and temperature for orientedly immobilized glucose oxidase are 4.0 and 57 ℃ respectively, and its Michaelis constant (Km) is 15.84 mmol/L. Compared with the free and the randomly immobilized glucose oxidases, the optimum pH of the orientedly immobilized glucose oxidase tends more acidic and its pH range is wider. Orientedly immobilized glucose oxidase also presents the higher temperature resistance and the better affinity to the substrate.

Key words: glucose oxidase, concanavalin A, oriented immobilization

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