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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.
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页码:208 / 215
页数:8
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