固定化酶反应器作为蛋白质组学研究中“bottom-up”策略重要的组件，具有酶解快速、酶解效率高、酶稳定性和活性高、简单易操作、能够与多种检测方式联用等优点，对于发展高效快速的蛋白质组学分析方法具有重要意义。本文就固定化酶反应器的制备方法及其在蛋白质组学中的应用做简单的概述，着重介绍酶的固定方法、固定化酶的载体、用于固定的酶的种类。近几年固定化酶反应器的研究集中于提高固酶量、保持酶活性、增加酶解效率、减小非特异性吸附等方面。研究结果表明，采用纳米材料、整体材料等新型载体，提高载体亲水性，采用多酶同时酶解等方法能够有效改善固定化酶反应器的性能，提高蛋白质的鉴定效率。
As an important part in“bottom-up”strategy of proteomics,immobilized enzyme reactors have great significance in the development of fast and more efficient protein analytical method,owing to its advantages of high speed and enzymatic efficiency,good stability and activity,easy operation,and the possibility of hyphenating with multiple detection instruments. In this paper,the preparation methods of immobilized enzyme reactors and their applications in proteomic investigation are introduced,focusing on the nature of enzymes,the immobilization methods and the carrier materials used for immobilizing enzyme. In recent years,the investiga-tions are focused on increasing the immobilization amounts of enzyme,keeping enzymatic activity,improving enzymatic efficiency and decreasing nonspecific adsorption. The investiga-tion results showed that by using novel carriers such as nanomaterial and monolith,increasing of hydrophilicity of carrier and tandem hydrolysis with multiple enzymes can greatly improve the performance of immobilized enzyme reactors and increase protein identification efficiencies.
参考文献
| [1] | Malvi, B.;Gupta, S.S..Encapsulation of enzyme in large mesoporous material with small mesoporous windows[J].Chemical communications,201263(63):7853-7855. |
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