Activity enhancement of CotA laccase by hydrophilic engineering, histidine tag optimization and static culture
文献类型：期刊论文
| 作者 | Li, Lei1,2,3; Xie, Tian2,3; Liu, Zhongchuan2,3; Feng, Hong1; Wang, Ganggang2,3 |
| 刊名 | PROTEIN ENGINEERING DESIGN & SELECTION
 |
| 出版日期 | 2018 |
| 卷号 | 31期号:1页码:1-5 |
| 关键词 | activity CotA laccase histidine tag hydrophobic residue static culture |
| ISSN号 | 1741-0126 |
| DOI | 10.1093/protein/gzx064 |
| 产权排序 | 2 |
| 文献子类 | Article |
| 英文摘要 | CotA protein from Bacillus subtilis is of laccase activity. The solubility of recombinant CotA is low, which hinders its application. In this study, histidine tag position optimization and hydrophilic engineering were applied to increase the yield and activity of CotA protein. The results showed that the protein yield of CotA with his tag at C-terminal (CH6-CotA) was four times of that of NH6-CotA (His tag at N-terminal). Then, 23 single mutants were constructed by substitutions of hydrophobic residues with hydrophilic amino acids. Among them, the protein yield of the mutant F207Y was increased by 30%; the catalytic activity (kcat/Km) of V403T and P455S was two and three times higher than that of CH6-CotA, respectively. Finally, triple mutant F2071Y/V403T/P455S with C-terminal his-tag (CH6-TSY) was constructed. When the proteins were expressed in microanaerobic condition, the activities of mutants CH6-P455S and CH6-TSY were enhanced about 48- and 42-folds compared to that of NH6-CotA in non-static culture. |
| 学科主题 | Biochemistry ; Biophysics |
| URL标识 | 查看原文 |
| WOS关键词 | ALKALI-STABLE LACCASE ; BACTERIAL LACCASE ; BACILLUS-SUBTILIS ; PICHIA-PASTORIS ; BIOCHEMICAL-CHARACTERIZATION ; DIRECTED EVOLUTION ; CRYSTAL-STRUCTURE ; ESCHERICHIA-COLI ; EXPRESSION ; LICHENIFORMIS |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 语种 | 英语 |
| WOS记录号 | WOS:000423199100001 |
| 出版者 | OXFORD UNIV PRESS |
| 源URL | [http://210.75.237.14/handle/351003/30366]  |
| 专题 | 环境治理与食品安全领域_应用与环境微生物研究 环境治理与食品安全领域_环境微生物四川省重点实验室 环境治理与食品安全领域_中国科学院环境与应用微生物重点实验室 |
| 作者单位 | 1.Sichuan Univ, Coll Life Sci, Key Lab Bioresources & Ecoenvironm, Sichuan Key Lab Mol Biol & Biotechnol, Chengdu 610064, Sichuan, Peoples R China; 2.Chinese Acad Sci, Chengdu Inst Biol, Key Lab Environm & Appl Microbiol, Chengdu 610041, Sichuan, Peoples R China; 3.Key Lab Environm Microbiol Sichuan Prov, Chengdu 610041, Sichuan, Peoples R China |
| 推荐引用方式 GB/T 7714 | Li, Lei,Xie, Tian,Liu, Zhongchuan,et al. Activity enhancement of CotA laccase by hydrophilic engineering, histidine tag optimization and static culture[J]. PROTEIN ENGINEERING DESIGN & SELECTION,2018,31(1):1-5. |
| APA | Li, Lei,Xie, Tian,Liu, Zhongchuan,Feng, Hong,&Wang, Ganggang.(2018).Activity enhancement of CotA laccase by hydrophilic engineering, histidine tag optimization and static culture.PROTEIN ENGINEERING DESIGN & SELECTION,31(1),1-5. |
| MLA | Li, Lei,et al."Activity enhancement of CotA laccase by hydrophilic engineering, histidine tag optimization and static culture".PROTEIN ENGINEERING DESIGN & SELECTION 31.1(2018):1-5. |
入库方式： OAI收割
来源：成都生物研究所
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