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Covalent immobilization of trypsin onto thermo-sensitive poly(N-isopropylacrylamide-co-acrylic acid) microspheres with high activity and stability
Lai, Enping1,2; Wang, Yuxia2; Wei, Yi2; Li, Guang1; Ma, Guanghui2
2016-06-05
Source PublicationJOURNAL OF APPLIED POLYMER SCIENCE
ISSN0021-8995
Volume133Issue:21Pages:43343
Abstract

Poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAM-co-AA)) microspheres with a high copolymerized AA content were fabricated using rapid membrane emulsification technique. The uniform size, good hydrophilicity, and thermo sensitivity of the microspheres were favorable for trypsin immobilization. Trypsin molecules were immobilized onto the microspheres surfaces by covalent attachment. The effects of various parameters such as immobilization pH value, enzyme concentration, concentration of buffer solution, and immobilization time on protein loading amount and enzyme activity were systematically investigated. Under the optimum conditions, the protein loading was 493 +/- 20 mgg(-1) and the activity yield of immobilized trypsin was 155%+/- 3%. The maximum activity (V-max) and Michaelis constant (K-m) of immobilized enzyme were found to be 0.74 Ms-1 and 0.54 mM, respectively. The immobilized trypsin showed better thermal and storage stability than the free trypsin. The enzyme-immobilized microspheres with high protein loading amount still can show a thermo reversible phase transition behavior. The research could provide a strategy to immobilize enzyme for application in proteomics. (c) 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 43343.

KeywordBioengineering Emulsion Polymerization Gels Stimuli-sensitive Polymers
SubtypeArticle
WOS HeadingsScience & Technology ; Physical Sciences
DOI10.1002/app.43343
URL查看原文
Indexed BySCI
Language英语
WOS KeywordSubsequent Suspension Polymerization ; Enzyme-immobilization ; Emulsification Technique ; Protein-analysis ; Temperature ; Proteomics ; Microgels ; Hydrogel ; Nanoparticles ; Purification
WOS Research AreaPolymer Science
WOS SubjectPolymer Science
Funding OrganizationNational Natural Science Foundation of China(51173187 ; 20706053 ; 21306208)
WOS IDWOS:000371249800001
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/20643
Collection研究所(批量导入)
Corresponding AuthorLi, Guang
Affiliation1.Donghua Univ, Coll Mat Sci & Engn, State Key Lab Modificat Chem Fibers & Polymer Mat, Shanghai 201620, Peoples R China
2.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China
Recommended Citation
GB/T 7714
Lai, Enping,Wang, Yuxia,Wei, Yi,et al. Covalent immobilization of trypsin onto thermo-sensitive poly(N-isopropylacrylamide-co-acrylic acid) microspheres with high activity and stability[J]. JOURNAL OF APPLIED POLYMER SCIENCE,2016,133(21):43343.
APA Lai, Enping,Wang, Yuxia,Wei, Yi,Li, Guang,&Ma, Guanghui.(2016).Covalent immobilization of trypsin onto thermo-sensitive poly(N-isopropylacrylamide-co-acrylic acid) microspheres with high activity and stability.JOURNAL OF APPLIED POLYMER SCIENCE,133(21),43343.
MLA Lai, Enping,et al."Covalent immobilization of trypsin onto thermo-sensitive poly(N-isopropylacrylamide-co-acrylic acid) microspheres with high activity and stability".JOURNAL OF APPLIED POLYMER SCIENCE 133.21(2016):43343.
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