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Stabilization of alpha-chymotrypsin by covalent immobilization on amine-functionalized superparamagnetic nanogel
Hong, J(洪军); Gong, PJ(龚培军); Xu, DM(徐冬梅); Dong, L; Yao, SD(姚思德); Yao, S (reprint author), Chinese Acad Sci, Shanghai Inst Appl Phys, Shanghai 201800, Peoples R China
2007
Source PublicationJOURNAL OF BIOTECHNOLOGY
ISSN0168-1656
Volume128Issue:3Pages:597
AbstractStabilization of alpha-chymotrypsin (CT) by covalent immobilization on the amine-functionalized magnetic nanogel was studied. The amino groups containing superparamagnetic nanogel was obtained by Hoffman degradation of the polyacrylamide (PAM)-coated Fe3O4 nanoparticles prepared by facile photochemical in situ polymerization. CT was then covalently bound to the magnetic nanogel with reactive amino groups by using 1-ethyl-3-(3-dimethylaminepropyl) carbodiimide as coupling reagent. The binding capacity was determined to be 61 mg enzyme/g nanogel by BCA protein assay. Specific activity of the immobilized CT was measured to be 0.93 U/(mg min), 59.3% as that of free CT. The obtained immobilized enzyme had better resistance to temperature and pH inactivation in comparison to free enzyme and thus widened the ranges of reaction pH and temperature. The immobilized enzyme exhibited good thermostability, storage stability and reusability. Kinetic parameters were determined for both the immobilized and free enzyme. The value of K,, of the immobilized enzyme was larger than did the free form, whereas the V-max was smaller for the immobilized enzyme. (c) 2006 Elsevier B.V. All rights reserved.
KeywordAmine-functionalized Superparamagnetic Nanogel Photochemical In Situ Polymerization Covalent Immobilization Alpha-chymotrypsin
Subject AreaPhysics
Indexed BySCI
Language英语
Funding Project应物所项目组
WOS IDWOS:000244404400016
Citation statistics
Cited Times:77[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.sinap.ac.cn/handle/331007/8194
Collection中科院上海应用物理研究所2004-2010年
Corresponding AuthorYao, S (reprint author), Chinese Acad Sci, Shanghai Inst Appl Phys, Shanghai 201800, Peoples R China
Recommended Citation
GB/T 7714
Hong, J,Gong, PJ,Xu, DM,et al. Stabilization of alpha-chymotrypsin by covalent immobilization on amine-functionalized superparamagnetic nanogel[J]. JOURNAL OF BIOTECHNOLOGY,2007,128(3):597.
APA Hong, J,Gong, PJ,Xu, DM,Dong, L,Yao, SD,&Yao, S .(2007).Stabilization of alpha-chymotrypsin by covalent immobilization on amine-functionalized superparamagnetic nanogel.JOURNAL OF BIOTECHNOLOGY,128(3),597.
MLA Hong, J,et al."Stabilization of alpha-chymotrypsin by covalent immobilization on amine-functionalized superparamagnetic nanogel".JOURNAL OF BIOTECHNOLOGY 128.3(2007):597.
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