Expression and characterization of two secreted His(6)-tagged endo-beta-1,4-glucanases from the mollusc Ampullaria crossean in Pichia pastoris
文献类型：期刊论文
| 作者 | Guo, R; Ding, M; Zhang, SL; Xu, GJ; Zhao, FK |
| 刊名 | ACTA BIOCHIMICA ET BIOPHYSICA SINICA
 |
| 出版日期 | 2008 |
| 卷号 | 40期号:5页码:419-425 |
| 关键词 | cellulase endoglucanase EG27I EG27II Ampullaria crossean Pichia pastoris |
| 通讯作者 | Zhao, FK (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Key Lab Prote, Shanghai 20031, Peoples R China.,fkzhao@sibs.ac.cn |
| 英文摘要 | Two endo-beta-1,4-glucanase cDNAs, eg27I and eg27II, from the mollusc Ampullaria crossean were expressed in Pichia pastoris cells. The secreted His(6)-tagged proteins were purified in a single chromatography step. The purified recombinant EG27I and EG27II showed enzymatic activity on carboxylmethyl cellulose sodium salt at 15.31 U/mg and 12.40 U/mg, respectively. The optimum pH levels of the recombinant EG27I and EG27II were 5.5 and 5.5-6.0, respectively, and the optimum temperatures were 50 degrees C and 50 degrees C-55 degrees C, respectively. The pH stability study revealed that both EG27I and EG27II showed their highest stability at pH 8.0. Analysis of their thermostability indicated that both EG27I and EG27II were relatively stable up to 40 degrees C. Site-directed mutagenesis of Asp43 and Asp153 of both EG27I and EG27II showed that the two Asp residues are critical for the enzymatic activity. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | MULBERRY LONGICORN BEETLE ; ENZYMATIC-ACTIVITY ; APRIONA-GERMARI ; CELLULASE GENE ; CDNA CLONING ; ENDOGENOUS CELLULASE ; MOLECULAR-CLONING ; ENDOGLUCANASE-V ; PURIFICATION ; TERMITE |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000255930600007 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/1378]  |
| 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 | Guo, R,Ding, M,Zhang, SL,et al. Expression and characterization of two secreted His(6)-tagged endo-beta-1,4-glucanases from the mollusc Ampullaria crossean in Pichia pastoris[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2008,40(5):419-425. |
| APA | Guo, R,Ding, M,Zhang, SL,Xu, GJ,&Zhao, FK.(2008).Expression and characterization of two secreted His(6)-tagged endo-beta-1,4-glucanases from the mollusc Ampullaria crossean in Pichia pastoris.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,40(5),419-425. |
| MLA | Guo, R,et al."Expression and characterization of two secreted His(6)-tagged endo-beta-1,4-glucanases from the mollusc Ampullaria crossean in Pichia pastoris".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 40.5(2008):419-425. |
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