Molecular cloning and functional analysis of a recombinant ribosome-inactivating protein (alpha-momorcharin) from Momordica charantia
文献类型：期刊论文
| 作者 | Wang, Shuzhen1; Zhang, Yubo1; Liu, Honggao2,3; He, Ying1; Yan, Junjie1; Wu, Zhihua1; Ding, Yi1 |
| 刊名 | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
 |
| 出版日期 | 2012-11-01 |
| 卷号 | 96期号:4页码:939-950 |
| 关键词 | Ribosome-inactivating proteins Alpha-momorcharin Heterologous expression N-glycosidase Antifungal activity |
| ISSN号 | 0175-7598 |
| DOI | 10.1007/s00253-012-3886-6 |
| 英文摘要 | Alpha-momorcharin (alpha-MC), a member of the ribosome-inactivating protein (RIP) family, has been used not only as antiviral, antimicrobial, and antitumor agents, but also as toxicant to protozoa, insects, and fungi. In this study, we expressed the protein in Escherichia coli Rosetta (DE3) pLysS strain and purified it by nickel-nitrilotriacetic acid affinity chromatography. A total of 85 mg of homogeneous protein was obtained from 1 l culture supernatant of Rosetta (DE3) pLysS, showing a high recovery rate of 73.9%. Protein activity assay indicated that alpha-MC had both N-glycosidase activity and DNA-nuclease activity, the former releasing RIP diagnostic RNA fragment (Endo's fragment) from rice rRNAs and the latter converting supercoiled circular DNA of plasmid pET-32a(+) into linear conformations in a concentration-dependent manner. Specially, we found that alpha-MC could inhibit the mycelial growth of Fusarium solani and Fusarium oxysporum with IC50 values of 6.23 and 4.15 mu M, respectively. Results of optical microscopy and transmission electron microscopy demonstrated that alpha-MC caused extensive septum formation, loss of integrity of the cell wall, separation of the cytoplasm from the cell wall, deformation of cells with irregular budding sites, and apoptosis in F. solani. Moreover, alpha-MC was active against Pseudomonas aeruginosa with an IC50 value of 0.59 mu M. The alpha-MC protein carries a high potential for the design of new antifungal drugs or the development of transgenic crops resistant to pathogens. |
| 资助项目 | Scientific Research Program of Wuhan Municipality, Hubei Province, People's Republic of China[200820422170] ; Fundamental Research Fund for the Central Universities[204-274388] |
| WOS研究方向 | Biotechnology & Applied Microbiology |
| 语种 | 英语 |
| WOS记录号 | WOS:000310222300010 |
| 出版者 | SPRINGER |
| 源URL | [http://202.127.146.157/handle/2RYDP1HH/3871]  |
| 专题 | 中国科学院武汉植物园 |
| 通讯作者 | Ding, Yi |
| 作者单位 | 1.Wuhan Univ, State Key Lab Hybrid Rice, Coll Life Sci, Wuhan 430072, Hubei Province, Peoples R China 2.Minist Water Resources, Inst Hydroecol, Wuhan, Hubei Province, Peoples R China 3.Chinese Acad Sci, Wuhan, Hubei Province, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang, Shuzhen,Zhang, Yubo,Liu, Honggao,et al. Molecular cloning and functional analysis of a recombinant ribosome-inactivating protein (alpha-momorcharin) from Momordica charantia[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2012,96(4):939-950. |
| APA | Wang, Shuzhen.,Zhang, Yubo.,Liu, Honggao.,He, Ying.,Yan, Junjie.,...&Ding, Yi.(2012).Molecular cloning and functional analysis of a recombinant ribosome-inactivating protein (alpha-momorcharin) from Momordica charantia.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,96(4),939-950. |
| MLA | Wang, Shuzhen,et al."Molecular cloning and functional analysis of a recombinant ribosome-inactivating protein (alpha-momorcharin) from Momordica charantia".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 96.4(2012):939-950. |
入库方式： OAI收割
来源：武汉植物园
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