high level expression, purification, and characterization of the shrimp antimicrobial peptide, ch-penaeidin, in pichia pastoris
文献类型：期刊论文
| 作者 | Li L ; Wang JX ; Zhao Xianfeng ; Kang CH ; Liu N ; Xiang JH ; Li FH ; Sueda S ; Kondo H |
| 刊名 | Protein Expression and Purification
 |
| 出版日期 | 2005 |
| 卷号 | 39期号:2页码:144-151 |
| 关键词 | Chinese shrimp (Fenneropenaeus chinensis) antimicrobial peptide Pichia pastoris recombinant expression penaeidin PENAEUS-VANNAMEI CLONING PSD1 |
| 通讯作者 | Wang, JX (通讯作者), Shandong Univ, Sch Life Sci, Jinan 250100, Peoples R China |
| 收录类别 | SCI |
| 公开日期 | 2010-08-25 |
| 附注 | Penaeidins, members of a new family of antimicrobial peptides constitutively produced and stored in the haemocytes of penaeid shrimp, display antimicrobial activity against bacteria, and fungi. Here, a DNA sequence encoding the mature Ch-penaeidin peptide was cloned into the pPIC9K vector and transformed into Pichia pastoris. The transformed cells were screened for multi-copy plasmids using increasing concentrations of G418. Positive colonies carrying chromosomal integrations of the Chp gene were identified by phenotype and PCR. When transformed cells were induced with methanol, SDS-PAGE and Western blotting revealed the production of a similar to6100 Da recombinant CHP (rCHP) expression product. Large scale expression revealed that rCHP was produced at 108 mg/L under optimal conditions in the highest Chp-producing P. pastoris clone. The antimicrobial activities of rCHP were studied by liquid phase analysis, which revealed that rCHP exhibited activities against some Gram-negative and Gram-positive bacteria, but had a relatively low activity against some fungi. Purification of rCHP by cation exchange chromatography and subsequent automated amino acid sequencing revealed the presence of four additional amino acids (YVEF) at the N-terminus that belonged to the cleaved fusion signal peptide; these residues may account for the observed decrease in antifungal activity. Together, these observations indicate that rCHP is an effective antimicrobial peptide that can be successfully produced at high levels in the yeast, and therefore may be a potential antimicrobial candidate for practical use. (C) 2004 Elsevier Inc. All rights reserved. |
| 源URL | [http://124.16.136.157/handle/311060/4968]  |
| 专题 | 软件研究所_信息安全国家重点实验室_期刊论文 |
| 推荐引用方式 GB/T 7714 | Li L,Wang JX,Zhao Xianfeng,et al. high level expression, purification, and characterization of the shrimp antimicrobial peptide, ch-penaeidin, in pichia pastoris[J]. Protein Expression and Purification,2005,39(2):144-151. |
| APA | Li L.,Wang JX.,Zhao Xianfeng.,Kang CH.,Liu N.,...&Kondo H.(2005).high level expression, purification, and characterization of the shrimp antimicrobial peptide, ch-penaeidin, in pichia pastoris.Protein Expression and Purification,39(2),144-151. |
| MLA | Li L,et al."high level expression, purification, and characterization of the shrimp antimicrobial peptide, ch-penaeidin, in pichia pastoris".Protein Expression and Purification 39.2(2005):144-151. |
入库方式： OAI收割
来源：软件研究所
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