Covalent immobilization of trypsin onto thermo-sensitive poly(N-isopropylacrylamide-co-acrylic acid) microspheres with high activity and stability

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	Covalent immobilization of trypsin onto thermo-sensitive poly(N-isopropylacrylamide-co-acrylic acid) microspheres with high activity and stability
	Lai, Enping 1,2; Wang, Yuxia 2; Wei, Yi 2; Li, Guang 1; Ma, Guanghui 2
刊名	JOURNAL OF APPLIED POLYMER SCIENCE
	2016-06-05
卷号	133 期号:21 页码:43343
关键词	bioengineering emulsion polymerization gels stimuli-sensitive polymers
ISSN号	0021-8995
通讯作者	Li, G (reprint author), Donghua Univ, Coll Mat Sci & Engn, State Key Lab Modificat Chem Fibers & Polymer Mat, Shanghai 201620, Peoples R China.
英文摘要	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.
WOS标题词	Science & Technology ; Physical Sciences
类目[WOS]	Polymer Science
研究领域[WOS]	Polymer Science
关键词[WOS]	SUBSEQUENT SUSPENSION POLYMERIZATION ; ENZYME-IMMOBILIZATION ; EMULSIFICATION TECHNIQUE ; PROTEIN-ANALYSIS ; TEMPERATURE ; PROTEOMICS ; MICROGELS ; HYDROGEL ; NANOPARTICLES ; PURIFICATION
收录类别	SCI
原文出处	WILEY-BLACKWELL
语种	英语
WOS记录号	WOS:000371249800001
内容类型	期刊论文
源URL	[http://ir.ipe.ac.cn/handle/122111/20643]
专题	过程工程研究所_研究所（批量导入）
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
推荐引用方式 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.