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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
发表期刊JOURNAL OF APPLIED POLYMER SCIENCE
ISSN0021-8995
卷号133期号:21页码:43343
摘要

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.

关键词Bioengineering Emulsion Polymerization Gels Stimuli-sensitive Polymers
文章类型Article
WOS标题词Science & Technology ; Physical Sciences
DOI10.1002/app.43343
URL查看原文
收录类别SCI
语种英语
关键词[WOS]Subsequent Suspension Polymerization ; Enzyme-immobilization ; Emulsification Technique ; Protein-analysis ; Temperature ; Proteomics ; Microgels ; Hydrogel ; Nanoparticles ; Purification
WOS研究方向Polymer Science
WOS类目Polymer Science
项目资助者National Natural Science Foundation of China(51173187 ; 20706053 ; 21306208)
WOS记录号WOS:000371249800001
引用统计
文献类型期刊论文
条目标识符http://ir.ipe.ac.cn/handle/122111/20643
专题研究所(批量导入)
通讯作者Li, Guang
作者单位1.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
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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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