Improving specific activity and thermostability of Escherichia coli phytase by structure-based rational design
文献类型：期刊论文
| 作者 | Wu, Tzu-Hui1; Chen, Chun-Chi3; Cheng, Ya-Shan4,5; Ko, Tzu-Ping6; Lin, Cheng-Yen4,5; Lai, Hui-Lin4,5; Huang, Ting-Yung4,5; Liu, Je-Ruei1,2,7; Guo, Rey-Ting3 |
| 刊名 | JOURNAL OF BIOTECHNOLOGY
 |
| 出版日期 | 2014-04-10 |
| 卷号 | 175期号:?页码:1-6 |
| 关键词 | Animal feed Phytate Protein engineering Sites-pecific mutagenesisa |
| 英文摘要 | Escherichia coli phytase (EcAppA) which hydrolyzes phytate has been widely applied in the feed industry, but the need to improve the enzyme activity and thermostability remains. Here, we conduct rational design with two strategies to enhance the EcAppA performance. First, residues near the substrate binding pocket of EcAppA were modified according to the consensus sequence of two highly active Citrobacter phywtases. One out of the eleven mutants, V89T, exhibited 17.5% increase in catalytic activity, whichmight be a result of stabilized protein folding. Second, the EcAppA glycosylation pattern was modifiedin accordance with the Citrobacter phytases. An N-glycosylation motif near the substrate binding sitewas disrupted to remove spatial hindrance for phytate entry and product departure. The de-glycosylatedmutants showed 9.6% increase in specific activity. On the other hand, the EcAppA mutants that adopt Nglycosylation motifs from CbAppA showed improved thermostability that three mutants carrying singleNglycosylation motif exhibited 5.6-9.5% residual activity after treatment at 80. C (1.8% for wild type). Furthermore, the mutant carrying all three glycosylation motifs exhibited 27% residual activity. In conclusion, a successful rational design was performed to obtain several useful EcAppA mutants with betterproperties for further applications. (C) 2014 Elsevier B. V. All rights reserved. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biotechnology & Applied Microbiology |
| 研究领域[WOS] | Biotechnology & Applied Microbiology |
| 关键词[WOS] | PICHIA-PASTORIS ; CRYSTAL-STRUCTURES ; GLYCOSYLATION ; EXPRESSION ; COMPLEX ; EFFICIENCY ; POULTRY ; CLONING ; FEED ; GENE |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000333090200001 |
| 源URL | [http://124.16.173.210/handle/834782/1354]  |
| 专题 | 天津工业生物技术研究所_结构生物学与蛋白酶学实验室 郭瑞庭_期刊论文 |
| 作者单位 | 1.Natl Taiwan Univ, Inst Biotechnol, Taipei 300308, Taiwan 2.Natl Taiwan Univ, Dept Anim Sci & Technol, Taipei 106, Taiwan 3.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Ind Enzymes Natl Engn Lab, Tianjin 300308, Peoples R China 4.Genozyme Biotechnol Inc, Taipei 106, Taiwan 5.AsiaPac Biotechnol Co Ltd, Dongguan 523808, Peoples R China 6.Acad Sinica, Inst Biol Chem, Taipei 115, Taiwan 7.Acad Sinica, Agr Biotechnol Res Ctr, Taipei 115, Taiwan |
| 推荐引用方式 GB/T 7714 | Wu, Tzu-Hui,Chen, Chun-Chi,Cheng, Ya-Shan,et al. Improving specific activity and thermostability of Escherichia coli phytase by structure-based rational design[J]. JOURNAL OF BIOTECHNOLOGY,2014,175(?):1-6. |
| APA | Wu, Tzu-Hui.,Chen, Chun-Chi.,Cheng, Ya-Shan.,Ko, Tzu-Ping.,Lin, Cheng-Yen.,...&Guo, Rey-Ting.(2014).Improving specific activity and thermostability of Escherichia coli phytase by structure-based rational design.JOURNAL OF BIOTECHNOLOGY,175(?),1-6. |
| MLA | Wu, Tzu-Hui,et al."Improving specific activity and thermostability of Escherichia coli phytase by structure-based rational design".JOURNAL OF BIOTECHNOLOGY 175.?(2014):1-6. |
入库方式： OAI收割
来源：天津工业生物技术研究所
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