基于共价键合法的毛细管固定化酶微反应器的制备

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英文篇名：Enzyme assay by capillary electrophoresis with an on-column immobilized enzyme microreactor created by a covalent binding technique 作者：张艳梅 ; 李凤 ; 赵晓燕 ; 赵志勇 ; 周昌艳 ; 康经武 英文作者：ZHANG Yan-mei;LI Feng;ZHAO Xiao-yan;ZHAO Zhi-yong;ZHOU Chang-yan;KANG Jing-wu;Institute for Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences;School of Chemical Engineering,Xi'an University;State Key Laboratory of Bioorganic and Natural Products Chemistry,Shanghai Institute of Organic Chemistry,Chinese Academy of Sciences; 关键词：毛细管固定化酶微反应器 ; 乙酰胆碱酯酶 ; 毛细管电泳 英文关键词：Capillary immobilized enzyme microreactor;;Acetylcholinesterase;;Capillary electrophoresis 中文刊名：FXSY 英文刊名：Chinese Journal of Analysis Laboratory 机构：上海市农业科学院农产品质量标准与检测技术研究所;西安文理学院化学工程学院;中国科学院上海有机化学研究所生命有机化学国家重点实验室; 出版日期：2019-03-21 16:48 出版单位：分析试验室 年：2019 期：v.38 基金：上海市市级农口系统青年人才成长计划资助项目(沪农青字(2018)第1-35号);; 国家食用菌产品质量安全风险评估专项(GJFP2018006);; 上海市农业科学院青年科技人员“助跑”计划资助 语种：中文; 页：FXSY201904010 页数：4 CN：04 ISSN：11-2017/TF 分类号：52-55

摘要

制备了一款开管式乙酰胆碱酯酶(ACHE)毛细管酶微反应器。该制备方法中,首先在毛细管内壁修饰上末端带有双键的硅烷化试剂,再采用N-丙烯酰氧琥珀酰亚胺作为功能单体,通过自由基聚合将其修饰在毛细管内壁上,最后通过共价键合的固定化方式,十分简便地实现了毛细管内壁的ACHE固定化。测试连续制备的6个ACHE酶微反应器,其产率的相对标准偏差(RSD)在3%以下;制备的ACHE酶微反应器在1 min孵育时间里的产率高达45%左右,可以较快进行酶活测试,每次分离仅需2. 5 min,10 h内重复使用40次后仍然能够保持酶活性,表明制备的ACHE酶微反应器具有较好的重复利用性和较高的酶活性。
        An open-tubular acetylcholinesterase( ACHE) immobilized enzymatic microreactor was prepared by the free radical polymerization with n-acryloyloxysuccinimide( NAS) as the organic functional monomer. The ACHE was covalently immobilized,and the ACHE reactor was easily prepared in two steps: firstly,fused-silica capillary was modified by 3-( trimethoxysilyl) propyl methacrylate to introduce C = C double bonds; subsequently,ACHE was covalently immobilized with NAS. The relative standard deviation( RSD,n = 6) of the yield of six ACHE enzyme microreactors was below 3%. 1 min incubation time led to the yield up to 45%. The substrate and product were separated by CE within 2. 5 minute,and the activity of the immobilized ACHE remained after 40 runs in 10 hours. The results demonstrated that the ACHE-immobilized capillary microreactor had better reusability and higher enzyme activity.

引文

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