Immobilization of horseradish peroxidase on chitosan for use in nonaqueous media

被引:40
作者
Bindhu, LV [1 ]
Abraham, ET [1 ]
机构
[1] CSIR, Reg Res Lab, Biochem Proc Div, Trivandrum 695019, Kerala, India
关键词
enzymes; catalysis; supports;
D O I
10.1002/app.11815
中图分类号
O63 [高分子化学(高聚物)];
学科分类号
070305 ; 080501 ; 081704 ;
摘要
Chitosan, a natural polysaccharide, was used for the covalent immobilization of horseradish peroxidase, an enzyme of high synthetic utility, with the carbodiimide method. Of the enzyme, 62% was immobilized on chitosan when 1-ethyl-3-(3-dimethylaminopropyl carbodiimide) was used as the peptide coupling agent. The influence of different parameters, such as the enzyme concentration, carbodiimide concentration, and incubation period, on the activity retention of the immobilized enzyme was investigated. Kinetic studies using horseradish peroxidase immobilized on chitosan revealed the effects of several parameters, such as the substrate hydrophilicity and hydrophobicity, the solubility of substrates in the medium, the solvent hydrophobicity, and the support aquaphilicity, on the catalytic activity of the immobilized enzyme in nonaqueous media. General rules for the optimization of solvents for nonaqueous enzymology based on the partitioning of the solvent were not applicable for the immobilized horseradish peroxidase. The catalytic efficiency was greatest when o-phenylene diamine was used as the substrate and least when guaiacol was used. The aquaphilicity of the support played an important role in the kinetics of the immobilized horseradish peroxidase in water-miscible solvents. The results were promising for the future development of chitosan-immobilized enzymes for use in organic media. (C) 2003 Wiley Periodicals, Inc.
引用
收藏
页码:1456 / 1464
页数:9
相关论文
共 49 条
[41]   Enzyme encapsulation on chitosan microbeads [J].
Siso, MIG ;
Lang, E ;
CarrenoGomez, B ;
Becerra, M ;
Espinar, FO ;
Mendez, JB .
PROCESS BIOCHEMISTRY, 1997, 32 (03) :211-216
[42]   FACTORS AFFECTING ENZYME CHARACTERISTICS OF BILIRUBIN OXIDASE SUSPENSIONS IN ORGANIC-SOLVENTS [J].
SKRIKAALEXOPOULOS, E ;
FREEDMAN, RB .
BIOTECHNOLOGY AND BIOENGINEERING, 1993, 41 (09) :887-893
[43]   A novel chitosan derivative to immobilize α-L-rhamnopyranosidase from Aspergillus niger for application in beverage technologies [J].
Spagna, G ;
Barbagallo, RN ;
Casarini, D ;
Pifferi, PG .
ENZYME AND MICROBIAL TECHNOLOGY, 2001, 28 (4-5) :427-438
[44]  
Vianello F, 2000, BIOTECHNOL BIOENG, V68, P488, DOI 10.1002/(SICI)1097-0290(20000605)68:5<488::AID-BIT2>3.0.CO
[45]  
2-Q
[46]   IMPROVED ACTIVITY RETENTION OF ENZYMES DEPOSITED ON SOLID SUPPORTS [J].
WEHTJE, E ;
ADLERCREUTZ, P ;
MATTIASSON, B .
BIOTECHNOLOGY AND BIOENGINEERING, 1993, 41 (02) :171-178
[47]   CARRIER-BOUND PROTEINS - PROPERTIES OF PEROXIDASE BOUND TO INSOLUBLE CARBOXYMETHYLCELLULOSE PARTICLES [J].
WELIKY, N ;
BROWN, FS ;
DALE, EC .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1969, 131 (01) :1-&
[48]  
Xu JJ, 1998, ELECTROANAL, V10, P713, DOI 10.1002/(SICI)1521-4109(199808)10:10<713::AID-ELAN713>3.0.CO
[49]  
2-6