The in vitro directed evolution of e-coli alkaline phosphatase
文献类型：期刊论文
| 作者 | Xu, HF; Zhang, XE; Zhang, ZP; Zhang, YM |
| 刊名 | Progress in biochemistry and biophysics
 |
| 出版日期 | 2003-02-01 |
| 卷号 | 30期号:1页码:89-94 |
| 关键词 | Directed evolution Error prone pcr E. coli alkaline phosphatase Catalytic activity |
| ISSN号 | 1000-3282 |
| 英文摘要 | The evolution of phoa gene fragment distant from the asp101-ser102-ala103 encoding region to increase the catalytic activity of eap with a single mutant d101s as parent was directed. through two cycles of error prone pcr, coupled with a sensitive screening method, an evolved variant 4-186 was obtained. its catalytic activity was 3-fold higher than that of d101s parent and 35-fold more active than wild-type ear the kinetic analysis indicated that the evolved enzyme exhibits a higher substrate binding ability and a higher catalytic efficiency than the d101s parent enzyme. dna sequence revealed that 4-186 contains two amino acid substitutions, k167r and s374c, both of which locate neither the substrate-binding sites nor the metal-binding sites of eap. |
| WOS关键词 | ACTIVE-SITE ; CATALYTIC ACTIVITY ; IN-VITRO ; MUTAGENESIS ; MECHANISM ; RESIDUES ; ENZYME |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biophysics |
| WOS类目 | Biochemistry & Molecular Biology ; Biophysics |
| 语种 | 英语 |
| WOS记录号 | WOS:000181216300017 |
| 出版者 | SCIENCE CHINA PRESS |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2376130 |
| 专题 | 武汉病毒研究所 |
| 作者单位 | 1.Chinese Acad Sci, Inst Microbiol, Beijing 100080, Peoples R China 2.Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China 3.Univ London Imperial Coll Sci Technol & Med, Dept Biochem, London SW7 2AY, England |
| 推荐引用方式 GB/T 7714 | Xu, HF,Zhang, XE,Zhang, ZP,et al. The in vitro directed evolution of e-coli alkaline phosphatase[J]. Progress in biochemistry and biophysics,2003,30(1):89-94. |
| APA | Xu, HF,Zhang, XE,Zhang, ZP,&Zhang, YM.(2003).The in vitro directed evolution of e-coli alkaline phosphatase.Progress in biochemistry and biophysics,30(1),89-94. |
| MLA | Xu, HF,et al."The in vitro directed evolution of e-coli alkaline phosphatase".Progress in biochemistry and biophysics 30.1(2003):89-94. |
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来源：武汉病毒研究所
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