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题名: Covalent immobilization of trypsin on glycidyl methacrylate-modified cellulose membrane as enzyme reactor
作者: Jiang, HH;  Zou, HF;  Wang, HL;  Ni, JY;  Zhang, Q
关键词: glycidyl methacrylate(GMA) -modified cellulose membrane ;  immobilized trypsin enzyme ;  biological reactor ;  degradation of protein ;  MALDI-TOF-MS ;  cytochrome C
刊名: CHEMICAL JOURNAL OF CHINESE UNIVERSITIES-CHINESE
发表日期: 2000-05-01
卷: 21, 期:5, 页:702-706
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Multidisciplinary
研究领域[WOS]: Chemistry
英文摘要: The membrane of the activated glycidyl methacrylate (GMA)-modified cellulose was prepared and packed into a column piece by piece to make a microreactor by immobilization of trypsin. The microreactor based on the membrane medium showed the advantages of being cheaper, mechanically strong and chemically stable. The activity of immobilized trypsin towards N-benzoyl-L-arginine ethyl ester(BAEE) was 17 800 U/g dry membrane, and was 52% of that for free enzyme. Besides, the effects of pH value of buffer, temperature, ionic strength, organic modifier, and protein denaturants on the activity of immobilized trypsin were investigated in comparison to the free trypsin, and thermal stability also was found especially to be improved after immobilization. The activity of the immobilized trypsin showed no decay after continuously pumping BAEE through immobilized trypsin microreactor for 24 h at 40 degrees C and 0. 5 mL/min. Finally, the cytochrome C was digested by microreactor and the products were analysed by MALDI-TOF-MS. The peptide mapping for a protein by combination of the immobilized enzyme membrane microreactor and MALDI-TOF-MS has been developed.
关键词[WOS]: PERFORMANCE LIQUID-CHROMATOGRAPHY ;  CAPILLARY ZONE ELECTROPHORESIS ;  IONIZATION MASS-SPECTROMETRY ;  TRYPTIC DIGEST ;  GROWTH-HORMONE ;  PURIFICATION ;  SEPARATION ;  PEPTIDE ;  PHASE ;  ANTIBODIES
语种: 英语
WOS记录号: WOS:000087161200011
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/138668
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Dalian 116011, Peoples R China

Recommended Citation:
Jiang, HH,Zou, HF,Wang, HL,et al. Covalent immobilization of trypsin on glycidyl methacrylate-modified cellulose membrane as enzyme reactor[J]. CHEMICAL JOURNAL OF CHINESE UNIVERSITIES-CHINESE,2000,21(5):702-706.
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