High expression of Trigonopsis variabilis D-amino acid oxidase in Pichia pastoris
文献类型：期刊论文
| 作者 | Yu, J; Li, DY; Zhang, YJ; Yang, S; Li, RB; Yuan, ZY |
| 刊名 | JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
 |
| 出版日期 | 2002 |
| 卷号 | 18期号:1页码:291-297 |
| 关键词 | D-amino acid oxidase Pichia pastoris cloning expression fermentation |
| 通讯作者 | Yuan, ZY (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China., |
| 英文摘要 | To explore a new approach of high expression Of D-amino acid oxidase (DAAO) in Pichia pastoris, a gene encoding DAAO from Trigonopsis variabilis (TvDAAO gene) deleted intron was prepared by PCR amplification and cloned into the intracellular expression vector pPIC3.5K. The expression plasmid pPIC3.5K-DAAO linearized by SalI was transformed into Pichia pastoris strain GS 115 (his(-)mut(+)). By means of MM and MD plates and PCR, the recombinant P pastoris strains (his(+)mut+) were obtained. Activity assay and SDS-PAGE demonstrated that DAAO was intracellularly expressed in R pastoris with the induction of methanol. The recombinant strain PD27 with the highest expression of DAAO was screened through activity assay and its high-density fermentation was carried out in a 1-1 fermentor. Activity assay and SDS-PAGE demonstrated that DAAO was intracellularly expressed in R pastoris with the induction of methanol. The recombinant cells with high expression of DAAO were screened and the high-density fermentation was carried out in a 1-1 fermentor. Interestingly, the DAAO expression level reached up to 473 U/g dry cell weight in fermentation yield. Finally, 1-hexanol was used to break recombinant cells and the specific activity of DAAO was 1.46 U/mg protein in crude extraction. (C) 2002 Elsevier Science B.V. All rights reserved. |
| 学科主题 | Biochemistry & Molecular Biology; Chemistry |
| 类目[WOS] | Biochemistry & Molecular Biology ; Chemistry, Physical |
| 关键词[WOS] | CEPHALOSPORIN-C ; CLONING ; CDNA |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000178982300014 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2560]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Yu, J,Li, DY,Zhang, YJ,et al. High expression of Trigonopsis variabilis D-amino acid oxidase in Pichia pastoris[J]. JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC,2002,18(1):291-297. |
| APA | Yu, J,Li, DY,Zhang, YJ,Yang, S,Li, RB,&Yuan, ZY.(2002).High expression of Trigonopsis variabilis D-amino acid oxidase in Pichia pastoris.JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC,18(1),291-297. |
| MLA | Yu, J,et al."High expression of Trigonopsis variabilis D-amino acid oxidase in Pichia pastoris".JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC 18.1(2002):291-297. |
入库方式： OAI收割
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。