Soluble Fusion Expression and Characterization of Human Beta-defensin 3 Using a Novel Approach
文献类型：期刊论文
| 作者 | Dong, Jing4; Yu, Heguo2; Zhang, Yonglian2; Diao, Hua2; Lin, Donghai1,3,4 |
| 刊名 | PROTEIN AND PEPTIDE LETTERS
 |
| 出版日期 | 2011-11 |
| 卷号 | 18期号:11页码:1126-1132 |
| 关键词 | Expression purification intein human beta-defensin3 antimicrobial activity |
| ISSN号 | 0929-8665 |
| DOI | 10.2174/092986611797201020 |
| 文献子类 | Article |
| 英文摘要 | Human beta-defensin 3 (DEFB103) is a recently identified small cysteine-rich cationic peptide expressed ubiquitously upon local microbial invasion. A number of accumulating evidences indicate that this peptide is involved in many biological processes, including microbicidal activities, chemoattraction, and immunomodulation. In this article, we describe a novel approach through which we performed the expression and purification of the recombinant DEFB103 peptide in Escherichia coli (E. coli) based on the pTWIN1 expression system. This approach does not introduce any extra residues to the peptide product, and also eliminates the requirement of removing the fusion tag by exogenous proteases. A high yield of 112 mg of soluble fusion DEFB103 was obtained in 1 liter of Luria-Bertani (LB) medium. By one-step affinity chromatography and on-column, auto-cleavage of the fusion tag, the mature DEFB103 peptide was produced with a yield of 30 mg/L LB. The purified DEFB103 peptide demonstrated strong antimicrobial activities against E. coli, S. aureus and C. albicans, which were representatives of Gram-negative and Gram-positive bacteria and fungi, respectively. Using this novel approach, we have successfully expressed and purified several human defensins with significant bioactivities. Our work may be helpful for structural and functional studies of other human defensins, and also for the production of human defensins. |
| WOS关键词 | HUMAN BETA-DEFENSIN-3 ; ESCHERICHIA-COLI ; PEPTIDES ; PROTEIN ; SYSTEM ; PURIFICATION ; LIGATION ; BACTERIA ; BINDING ; INTEIN |
| 资助项目 | Program of Shanghai Subject Chief Scientist[09XD1405100] |
| WOS研究方向 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:000298538600009 |
| 出版者 | BENTHAM SCIENCE PUBL LTD |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/278344]  |
| 专题 | 分析化学研究室 |
| 通讯作者 | Diao, Hua |
| 作者单位 | 1.Xiamen Univ, Key Lab Chem Biol Fujian Prov, Coll Chem & Chem Engn, Xiamen 361005, Peoples R China; 2.Shanghai Inst Planned Parenthood Res, NPFPC Key Lab Contracept & Devices, Shanghai 200032, Peoples R China; 3.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China 4.China Pharmaceut Univ, Sch Life Sci & Technol, Nanjing 210009, Peoples R China; |
| 推荐引用方式 GB/T 7714 | Dong, Jing,Yu, Heguo,Zhang, Yonglian,et al. Soluble Fusion Expression and Characterization of Human Beta-defensin 3 Using a Novel Approach[J]. PROTEIN AND PEPTIDE LETTERS,2011,18(11):1126-1132. |
| APA | Dong, Jing,Yu, Heguo,Zhang, Yonglian,Diao, Hua,&Lin, Donghai.(2011).Soluble Fusion Expression and Characterization of Human Beta-defensin 3 Using a Novel Approach.PROTEIN AND PEPTIDE LETTERS,18(11),1126-1132. |
| MLA | Dong, Jing,et al."Soluble Fusion Expression and Characterization of Human Beta-defensin 3 Using a Novel Approach".PROTEIN AND PEPTIDE LETTERS 18.11(2011):1126-1132. |
入库方式： OAI收割
来源：上海药物研究所
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