On-line characterization of the activity and reaction kinetics of immobilized enzyme by high-performance frontal analysis
文献类型：期刊论文
| 作者 | Jiang, HH; Zou, HF; Wang, HL; Ni, JY; Zhang, Q; Zhang, YK |
| 刊名 | journal of chromatography a
 |
| 出版日期 | 2000-12-15 |
| 卷号 | 903期号:1-2页码:77-84 |
| 关键词 | immobilized trypsin reactor glycidyl methacrylate-modified membrane reaction kinetics frontal analysis enzymes |
| 英文摘要 | a microreactor by immobilized trypsin on the activated glycidyl methacrylate-modified cellulose membrane packed column was constructed, immobilized trypsin mirrored the properties of the free enzyme and showed high stability. a novel method to characterize the activity and reaction kinetics of the immobilized enzyme has been developed based on the frontal analysis of enzymatic reaction products, which was performed by the on-line monitoring of the absorption at 410 nm of p-nitroaniline from the hydrolysis of n-alpha -benzoyl-dl-arginine-p-nitroanilide (bapna). the hydrolytic activity of the immobilized enzyme was 55.6% of free trypsin. the apparent michaelis-menten kinetics constant (k-m) and v-max values measured by the frontal analysis method were, respectively, 0.12 mm and 0.079 mm min(-1) mg enzyme(-1). the former is very close to that observed by the static and off-line detection methods, but the latter is about 15% higher than that of the static method. inhibition of the immobilized trypsin by addition of benzamidine into substrate solution has been studied by the frontal analysis method. the apparent michaelis-menten constant of bapna (k-m), the inhibition constant of benzamidine (k-m) and v-max were determined. it was indicated that the interaction of bapna and benzamidine with trypsin is competitive, the k-m value was affected but the v-max was unaffected by the benzamidine concentration. (c) 2000 elsevier science b.v. all rights reserved. |
| WOS标题词 | science & technology ; life sciences & biomedicine ; physical sciences |
| 类目[WOS] | biochemical research methods ; chemistry, analytical |
| 研究领域[WOS] | biochemistry & molecular biology ; chemistry |
| 关键词[WOS] | affinity-chromatography ; glucose-oxidase ; purification ; membranes ; retention ; trypsin ; spacer |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000165977800008 |
| 公开日期 | 2015-11-10 |
| 源URL | [http://159.226.238.44/handle/321008/138773]  |
| 专题 | 大连化学物理研究所_中国科学院大连化学物理研究所 |
| 作者单位 | Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromat R&A Ctr, Dalian 116011, Peoples R China |
| 推荐引用方式 GB/T 7714 | Jiang, HH,Zou, HF,Wang, HL,et al. On-line characterization of the activity and reaction kinetics of immobilized enzyme by high-performance frontal analysis[J]. journal of chromatography a,2000,903(1-2):77-84. |
| APA | Jiang, HH,Zou, HF,Wang, HL,Ni, JY,Zhang, Q,&Zhang, YK.(2000).On-line characterization of the activity and reaction kinetics of immobilized enzyme by high-performance frontal analysis.journal of chromatography a,903(1-2),77-84. |
| MLA | Jiang, HH,et al."On-line characterization of the activity and reaction kinetics of immobilized enzyme by high-performance frontal analysis".journal of chromatography a 903.1-2(2000):77-84. |
入库方式： OAI收割
来源：大连化学物理研究所
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