Preparation and properties of thermoresponsive bioconjugates of trypsin

被引:3
作者
Raghava, Smita
Mondal, Kalyani
Gupta, Munishwar N. [1 ]
Pareek, Pradeep
Kuckling, Dirk
机构
[1] Indian Inst Technol, Dept Chem, New Delhi 110016, India
[2] Tech Univ Dresden, Inst Macromol Chem & Text Chem, Dresden, Germany
来源
ARTIFICIAL CELLS BLOOD SUBSTITUTES AND BIOTECHNOLOGY | 2006年 / 34卷 / 03期
关键词
enzyme immobilization; smart bioconjugates; trypsin; poly(N-isopropylacrylamide); smart polymers;
D O I
10.1080/10731190600683902
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Covalent attachment of enzymes and other proteins to the smart polymer, poly(N-isopropylacrylamide) [poly (NIPAAm)], has been widely used as a method for the preparation of thermosensitive protein conjugates. In the present study, reversible soluble-insoluble polymer-enzyme conjugates were prepared by conjugating a copolymer of NIPAAm with 5-mol % of 6-acrylaminohexanoic acid to trypsin by the carbodiimide-NHS (N-hydroxysuccinimide) coupling method. Four bioconjugates with different units of enzyme coupled to the matrix were prepared. Increased enzymatic activity in terms of high effectiveness factor (in the range of 3-5) was found in the conjugates. Kinetic parameters for the immobilized and free enzyme were determined. The V-max/K-m value of the enzyme significantly increased on immobilization by the factors in the range of 12-28. The immobilized enzyme also showed stability to autolysis at 50 degrees C.
引用
收藏
页码:323 / 336
页数:14
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