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浙江大学学报(工学版)
J4  2013, Vol. 47 Issue (1): 182-187    DOI: 10.3785/j.issn.1008-973X.2013.01.026
化学工程     
辣根过氧化物酶的新型包埋及活性研究
韩哲勇1, 厉瑾1, 宋锡瑾1, 王杰2
1. 浙江大学 化学工程与生物工程学系,浙江 杭州 310027；2. 浙江大学 化学系,浙江 杭州 310027
Novel encapsulation of horseradish peroxidase and its activity
HAN Zhe-yong1, LI Jin1, SONG Xi-jin1, WANG Jie2
1. Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China;
2. Department of Chemistry, Zhejiang University, Hangzhou 310027, China
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摘要：

N-丙烯酰氧基琥珀酰亚胺对辣根过氧化物酶（HRP）进行化学修饰,在酶分子表面接上不饱和双键,以聚乙二醇400二丙烯酸酯（单体A）、聚乙二醇400二甲基丙烯酸酯（单体B）、聚乙二醇200二丙烯酸酯（单体C）、聚乙二醇200二甲基丙烯酸酯（单体D）4种单体中的一种和丙烯酰胺为聚合原料,在引发剂作用下进行自由基原位聚合,使酶分子的表面形成多点连接的网状薄壳结构,获得4种新型包埋的辣根过氧化物酶.研究结果表明,聚乙二醇200（400）二丙烯酸酯包埋酶的活性和稳定性较高.在pH 为3.0、11.0和温度为70 ℃ 的条件下,CHRP的相对活性达到47.9%、39.7%、76.5%,重复使用5次后,C-HRP保留了62.1%的初始活性.在变性剂SDS和脲中浸泡30 d后,4种包埋酶的活性可以保持初始活性的36％以上,说明采用该新型包埋方法可以获得较高的酶活性和稳定性.
Abstract:

Horseradish peroxidase (HRP) was chemically modified to get unsaturated double bonds at the surface by N-acryloxysuccinimide. The free radical polymerization was initiated in a copolymer system, which contained acrylamide and one of four kinds of monomers, PEG400-diacrylate ester (A), PEG400-dimethacrylate ester (B), PEG200diacrylate ester (C) and PEG200dimethacrylate ester (D). Four novel encapsulated HRP were obtained and there were multipoint connected network and thin shell structure at the surface of enzymes. Results show that the activity of encapsulated enzymes with PEG200(400-diacrylate ester remains high and the stability is enhanced. C-HRP retains 47.9%, 39.7%, 76.5% of its relative activity under conditions of pH 3.0,11.00 and 70 ℃, respectively. When C-HRP was reused by five times, C-HRP was retained 62.1% of its initial activities. When encapsulated HRP was soaked for 30 days in SDS and urea solution, the relative activities of four encapsulated HRP were 36% of their initial activities, which showed that the novel encapsulated method can retain higher activity and stability．
出版日期: 2013-01-01
:  Q 55  
基金资助:

国家自然科学基金资助项目(20276067).
通讯作者: 宋锡瑾，女，副教授.     E-mail: xjsong@zju.edu.cn
作者简介: 韩哲勇（1987-）,男,硕士生,从事酶工程的研究.E-mail: 21028110@zju.edu.cn
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引用本文:

韩哲勇, 厉瑾, 宋锡瑾, 王杰. 辣根过氧化物酶的新型包埋及活性研究[J]. J4, 2013, 47(1): 182-187.

HAN Zhe-yong, LI Jin, SONG Xi-jin, WANG Jie. Novel encapsulation of horseradish peroxidase and its activity. J4, 2013, 47(1): 182-187.

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http://www.zjujournals.com/eng/CN/10.3785/j.issn.1008-973X.2013.01.026    或    http://www.zjujournals.com/eng/CN/Y2013/V47/I1/182

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