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›› 2012, Vol. 12 ›› Issue (5): 828-833.

• 生化工程专栏 • 上一篇    下一篇

阴离子型双功能含环氧基团酶固定化载体的制备及其固定辣根过氧化酶

雷青娟 高保娇 张正国   

  1. 中北大学化学工程系 中北大学化工系 中北大学化工系
  • 收稿日期:2012-07-17 修回日期:2012-08-01 出版日期:2012-10-20 发布日期:2012-10-20
  • 通讯作者: 雷青娟

Preparation of Anion-type Bifunctional Carrier with Epoxy Group and Its Immobilization of Horseradish Peroxidase

LEI Qing-juan GAO Bao-jiao ZHANG Zheng-guo   

  1. Department of Chemical Engineering, North University of China Department of Chemical Engineering, North University Department of Chemical Engineering, North University
  • Received:2012-07-17 Revised:2012-08-01 Online:2012-10-20 Published:2012-10-20
  • Contact: LEI Qing-juan

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被引次数

1

摘要： 利用接枝聚合在微米级硅胶微粒表面的甲基丙烯酸缩水甘油酯(GMA)和对羟基苯磺酸钠(SHBS)发生环氧基团的部分开环反应，制得阴离子型双功能复合载体SHBS-PGMA/SiO2，研究了辣根过氧化酶(HRP)与载体SHBS-PGMA/SiO2间的静电相互作用在共价固定HRP过程中的作用及其机理. 结果表明，在较大的pH值范围内，复合载体SHBS-PGMA/SiO2表面携带高密度负电荷，pH=6.0时HRP分子荷正电，酶蛋白分子与载体间会产生强静电相互作用，显著促进HRP的固定化；当载体表面SHBS的键合率约为18%时，静电相互作用对固酶的促进作用最强，固定化酶的偶联率和比活力最高. 离子强度对酶的固定化也有很大影响，加入NaCl对载体与酶蛋白之间的静电相互作用力产生屏蔽作用，增大NaCl浓度可使固定化酶的偶联率和比活力降低.

关键词: 辣根过氧化酶, 固定化酶, 甲基丙烯酸缩水甘油酯, 对羟基苯磺酸钠, 静电相互作用

Abstract: Glycidyl methacrylate (GMA) was graft-polymerized on micron-sized silica gel particles, obtaining the grafted particles PGMA/SiO2. The partial chain units of grafted macromolecule PGMA to produce ring-opening reaction of the epoxy groups with sodium 4-hydroxybenzenesulfonate (SHBS) as reagent were made, obtaining anion-type bifunctional composite carrier containing epoxy group, SHBS-PGMA/SiO2. With SHBS-PGMA/SiO2 as carrier, the immobilization of horseradish peroxidase (HRP) was carried out. The effect and mechanism of electrostatic interaction between the enzymic protein HRP and functional carrier on the enzyme immobilization were studied. The results show that the sulfonate anions on SHBS-PGMA/SiO2 particles enable the carrier particles to be negatively charged. As the pH value of the medium is 6.0, HRP molecules are positively charged, and the strong electrostatic interaction between the enzyme protein and the carrier will happen, which leads to strong physical adsorption of HRP on the carrier, and greatly promotes the chemical immobilization of HRP. For the carrier with an SHBS bonding rate of about 18%, there is the strongest electrostatic interaction between them, and the immobilized enzyme has the highest coupling rate and specific activity. The ionic strength affects HRP immobilization greatly. As electrolyte NaCl is added into the medium, the electrostatic interaction between them will be shielded, which makes the coupling rate and specific activity of immobilized enzyme decrease.

Key words: horseradish peroxidase, immobilized enzyme, glycidyl methacrylate, sodium 4-hydroxybenzenesulfonate, electrostatic interaction

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