收稿日期: 2010-12-26
修回日期: 2011-03-28
网络出版日期: 2011-08-02
基金资助
国家"973”计划项目( 2010BC732206) ; 国家自然科学基金资助项目( 30771689, 30972324) ; 教育部高校博士点基金资助项目( 20090172110022)
Immobilization of Laccase via Layer-by-Layer Self-Assembly and Its Enzymatic Properties
Received date: 2010-12-26
Revised date: 2011-03-28
Online published: 2011-08-02
Supported by
国家"973”计划项目( 2010BC732206) ; 国家自然科学基金资助项目( 30771689, 30972324) ; 教育部高校博士点基金资助项目( 20090172110022)
李辉 付时雨 彭林才 詹怀宇 . 漆酶的层层自组装固定化及其酶学性质[J]. 华南理工大学学报(自然科学版), 2011 , 39(9) : 158 -164 . DOI: 10.3969/j.issn.1000-565X.2011.09.027
Laccase was immobilized through the electrostatic layer-by-layer self-assembly,and cationic polyacrylamide ( CPAM) /laccase self-assembled films were prepared on solid substrate ( mica) . Then,the activities and morphologies of the films were characterized by means of the UV-Vis spectroscopy and the atomic force microscopy,and the enzymatic properties of the immobilized laccase were also analyzed. The results show that the activity of the
CPAM /laccase self-assembled film linearly increases with the number of layers,and the enzyme loading can be effectively controlled by changing the adsorption cycles,and that the laccase molecules are arranged orderly in the films,and the average height and RMS roughness of the self-assembled film increase with the number of layers.Moreover,it is found that the optimal pH value and temperature of the immobilized laccase are respectively 5. 0 and 50℃,which is 5 ℃ higher than that of free laccase,and the corresponding Michaelis constant for ABTS is 0. 042mmol /L,that the thermal stabilities of the immobilized laccase is significantly higher than those of free laccase,and that the immobilized laccase is of excellent reusability and high storage stability.
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