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http://dx.doi.org/10.4014/jmb.1304.03103

Chemically Modified Sepharose as Support for the Immobilization of Cholesterol Oxidase  

Yang, Hailin (The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University)
Chen, Yi (The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University)
Xin, Yu (The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University)
Zhang, Ling (The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University)
Zhang, Yuran (The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University)
Wang, Wu (The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University)
Publication Information
Journal of Microbiology and Biotechnology / v.23, no.9, 2013 , pp. 1212-1220 More about this Journal
Abstract
Because the cholesterol oxidase from Brevibacterium sp. M201008 was not as stable as the free enzyme form, it had been covalently immobilized onto chemically modified Sepharose particles via N-ethyl-N'-3-dimethylaminopropyl carbodiimide. The optimum immobilization conditions were determined, and the immobilized enzyme activity obtained was 12.01 U/g Sepharose-ethylenediamine. The immobilization of the enzyme was characterized by Fourier transform infrared spectroscopy. The immobilized enzyme exhibited the maximal activity at $35^{\circ}C$ and pH 7.5, which was unchanged compared with the free form. After being repeatedly used 20 times, the immobilized enzyme retained more than 40.43% of its original activity. The immobilized enzyme showed better operational stability, including wider thermal and pH ranges, and retained 62.87% activity after 20 days of storage at $4^{\circ}C$, which was longer than the free enzyme.
Keywords
Chemical modification; sepharose; cholesterol oxidase; immobilization; enzyme stability;
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