Purification of alpha-sarcin and an antifungal protein from mold (Aspergillus giganteus) by chitin affinity chromatography
文献类型：期刊论文
| 作者 | Liu, RS; Huang, H; Yang, Q; Liu, WY |
| 刊名 | PROTEIN EXPRESSION AND PURIFICATION
 |
| 出版日期 | 2002 |
| 卷号 | 25期号:1页码:50-58 |
| 关键词 | antifungal protein (AFP) Aspergillus giganteus MDH 18894 N-bromosuccinimide chitin affinity column chromatography glycochitin alpha-sarcin |
| 通讯作者 | Liu, WY (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Mol Biol, 320 Yue Yang Rd, Shanghai 200031, Peoples R China., |
| 英文摘要 | A simple method for preparation of alpha-sarcin and an antifungal protein (AFP) from mold (Aspergillus giganteus MDH 18894) has been developed. alpha-Sarcin and AFP were purified simultaneously by chitin affinity column chromatography and gel filtration. By this method, 4.5 mg of pure alpha-sarcin and 6.9 mg of pure AFP were obtained from 2 liters of culture medium. Compared with other purification methods such as ion-exchange column chromatography, this procedure was very simple and specific. The purified alpha-sarcin and AFP were homogeneous as characterized by SDS-polyacrylamide gel electrophoresis. Both alpha-sarcin and AFP exhibited the binding activity to generated chitin. Soluble glycochitin decreased the intensity of fluorescence of alpha-sarcin and made the lambda(em)m shift from 340 to 347 nm. Titration of alpha-sarcin with N-bromosuccinimide under native conditions revealed that two tryptophans (Trps) were all located in the core part of alpha-sarcin molecule. This indicated that Trps were not involved in the binding of alpha-sarcin to chitin. Glycochitin in the culture medium increased the expression of alpha-sarcin, while it had no effect on the expression of AFP. Unlike other ligands such as Cibacron blue for the affinity purification of alpha-sarcin and AFP, glycochitin increased the nuclease activity of alpha-sarcin. (C) 2002 Elsevier Science (USA). |
| 学科主题 | Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology |
| 类目[WOS] | Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 关键词[WOS] | RIBOSOME-INACTIVATING PROTEIN ; BINDING PROTEIN ; TRYPTOPHAN RESIDUES ; MECHANISM ; ASSAY |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000176592400007 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2410]  |
| 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 | Liu, RS,Huang, H,Yang, Q,et al. Purification of alpha-sarcin and an antifungal protein from mold (Aspergillus giganteus) by chitin affinity chromatography[J]. PROTEIN EXPRESSION AND PURIFICATION,2002,25(1):50-58. |
| APA | Liu, RS,Huang, H,Yang, Q,&Liu, WY.(2002).Purification of alpha-sarcin and an antifungal protein from mold (Aspergillus giganteus) by chitin affinity chromatography.PROTEIN EXPRESSION AND PURIFICATION,25(1),50-58. |
| MLA | Liu, RS,et al."Purification of alpha-sarcin and an antifungal protein from mold (Aspergillus giganteus) by chitin affinity chromatography".PROTEIN EXPRESSION AND PURIFICATION 25.1(2002):50-58. |
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