次价键力在化学键合法固定辣根过氧化酶过程中的作用机理

引用本文: 雷青娟, 高保娇, 张正国. 次价键力在化学键合法固定辣根过氧化酶过程中的作用机理[J]. 物理化学学报, 2011, 27(11): 2697-2704. doi: 10.3866/PKU.WHXB20111122 shu

Citation: LEI Qing-Juan, GAO Bao-Jiao, ZHANG Zheng-Guo. Action Mechanism of Secondary Bond Forces on Chemical Immobilization of Horseradish Peroxidase[J]. Acta Physico-Chimica Sinica, 2011, 27(11): 2697-2704. doi: 10.3866/PKU.WHXB20111122 shu

次价键力在化学键合法固定辣根过氧化酶过程中的作用机理

基金项目:
山西省自然科学基金(201002100843)资助项目 (201002100843)

摘要: 将甲基丙烯酸缩水甘油酯(GMA)接枝聚合在微米级硅胶微粒表面, 又通过环氧基团的开环反应将乙二胺(EDA)键合在接枝微粒表面, 制备了双功能复合载体EDA-PGMA/SiO₂, 通过共价偶联法实施了辣根过氧化酶(HRP)的固定化. 本文重点考察静电相互作用与疏水相互作用两种次价键力在共价键合法固定HRP过程中的作用, 探讨作用机理. 结果表明, 在水介质中, 复合载体EDA-PGMA/SiO₂表面胺基的质子化作用, 使载体微粒的ζ电位在较大的pH范围内保持正值; 当介质的pH=8.5, 大于HRP的等电点时, 酶蛋白与载体之间所产生的强静电相互作用会显著促进HRP的固定化; EDA的键合率在30%附近的载体, 静电相互作用对固酶的促进作用最强, 固定化酶的偶联率与比活力具有最高值. 疏水相互作用对化学法固定辣根过氧化酶也会产生明显的作用, 当以接枝微粒PGMA/SiO₂为载体时, 增大NaCl浓度, 可有效促进酶蛋白与载体之间的疏水相互作用, 提高固定化酶的偶联率与比活力.

关键词:

English

Action Mechanism of Secondary Bond Forces on Chemical Immobilization of Horseradish Peroxidase

1.
Department of Chemical Engineering, North University of China, Taiyuan 030051, P. R. China
Received Date: 27 June 2011
Available Online: 27 October 2011
Fund Project:
山西省自然科学基金(201002100843)资助项目

Abstract: Glycidyl methacrylate (GMA) was graft-polymerized onto micron-sized silica gel particles. Ethylenediamine (EDA) was bonded to the surfaces of PGMA/SiO₂ particles by the ring-opening reaction of epoxy groups resulting in the difunctional composite carrier, EDA-PGMA/SiO₂, which was used for enzyme immobilization. We immobilized horseradish peroxidase (HRP) using the chemical bonding method. In this work, the effects and action mechanisms of two secondary bond forces, electrostatic interaction and hydrophobic interaction, on the enzyme immobilization were investigated. The experimental results show that the protonated amino groups on the EDA-PGMA/SiO₂ particles enable the carrier particles to be positively charged and the zeta potential of the carrier particles are positive over a wider range of pH values. At a pH value of 8.5 for the medium, which is higher than the isoelectric point of HRP, the strong electrostatic interaction between the enzyme protein and the carrier significantly promotes the immobilization of HRP. For the carrier with an EDA bonding rate of about 30%, the strongest electrostatic interaction was observed between the enzyme protein and the carrier while the immobilized enzyme has the highest coupling rate and specific activity. Hydrophobic interaction between the enzyme protein and the carrier also affects HRP immobilization greatly. As the grafted particles PGMA/SiO₂ are used as the carrier, the addition of NaCl electrolyte will facilitate the hydrophobic interaction between the enzyme protein and the carrier and it will result in an increase in the coupling rate and specific activity of the immobilized enzyme.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

次价键力在化学键合法固定辣根过氧化酶过程中的作用机理

English

Action Mechanism of Secondary Bond Forces on Chemical Immobilization of Horseradish Peroxidase

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

次价键力在化学键合法固定辣根过氧化酶过程中的作用机理

English

Action Mechanism of Secondary Bond Forces on Chemical Immobilization of Horseradish Peroxidase

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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