Novel and efficient method for immobilization and stabilization of β-D-galactosidase by covalent attachment onto magnetic Fe3O4-chitosan nanoparticles

被引:229
作者
Pan, Chenliang [1 ]
Hu, Bing [1 ]
Li, Wei [1 ]
Sun, Yi [1 ]
Ye, Hong [1 ]
Zeng, Xiaoxiong [1 ]
机构
[1] Nanjing Agr Univ, Coll Food Sci & Technol, Nanjing 210095, Peoples R China
关键词
Magnetic Fe3O4 nanoparticle; Chitosan; beta-D-Galactosidase; Immobilization; Galactooligosaccharide; GLUTARALDEHYDE CROSS-LINKING; CANDIDA-RUGOSA LIPASE; LACTOSE HYDROLYSIS; GLUCOSE-OXIDASE; CHITOSAN; SUPPORT; BEADS; ACTIVATION; PARTICLES; INACTIVATION;
D O I
10.1016/j.molcatb.2009.07.003
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A novel and efficient immobilization of beta-D-galactosidase from Aspergillus oryzae has been developed by using magnetic Fe3O4-chitosan (Fe3O4-CS) nanoparticles as support. The magnetic Fe3O4-CS nanoparticles were prepared by electrostatic adsorption of chitosan onto the surface of Fe3O4 nanoparticles made through co-precipitation of Fe2+ and Fe3+. The resultant material was characterized by transmission electron microscopy. X-ray diffraction, Fourier transform infrared spectroscopy, vibrating sample magnetometry and thermogravimetric analysis. beta-D-Galactosidase was covalently immobilized onto the nanocomposites using glutaraldehyde as activating agent. The immobilization process was optimized by examining immobilized time, cross-linking time, enzyme concentration, glutaraldehyde concentration, the initial pH values of glutaraldehyde and the enzyme solution. As a result. the immobilized enzyme presented a higher storage, pH and thermal stability than the soluble enzyme. Calactooligosaccharide was formed with lactose as substrate by using the immobilized enzyme as biocatalyst, and a maximum yield of 15.5% (w/v) was achieved when about 50% lactose was hydrolyzed. Hence, the magnetic Fe3O4-chitosan nanoparticles are proved to be an effective support for the immobilization of beta-D-galactosidase. (C) 2009 Elsevier B.V. All rights reserved.
引用
收藏
页码:208 / 215
页数:8
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