Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli
文献类型：CNKI期刊论文
| 作者 | Jiang S(姜珊); Liu M(刘梅); Wang BJ(王宝杰); Jiang KY(蒋克勇); Wang L(王雷) |
| 发表日期 | 2011-11-15 |
| 出处 | Chinese Journal of Oceanology and Limnology
 |
| 关键词 | a nti-lipopolysaccharide factor optimization medium composition culture conditions statistical methodology |
| 英文摘要 | Anti-lipopolysaccharide factors (ALFs) are important antimicrobial peptides that are isolated from some aquatic species. In a previous study, we isolated ALF genes from Chinese mitten crab, Eriocheir sinensis. In this study, we optimized the production of a recombinant ALF by expressing E. sinensis ALF genes in Escherichia coli maintained in shake-flasks. In particular, we focused on optimization of both the medium composition and the culture condition. Various medium components were analyzed by the Plackett-Burman design, and two significant screened factors, (NH4)2SO4 and KH2PO4, were further optimized via the central composite design (CCD). Based on the CCD analysis, we investigated the induction start-up time, the isopropylthio-D-galactoside (IPTG) concentration, the post-induction time, and the temperature by response surface methodology. We found that the highest level of ALF fusion protein was achieved in the medium containing 1.89 g/L (NH4)2SO4 and 3.18 g/L KH2PO4, with a cell optical density of 0.8 at 600 nm before induction, an IPTG concentration of 0.5 mmol/L, a post-induction temperature of 32.7°C, and a post-induction time of 4 h. Applying the whole optimization strategy using all optimal factors improved the target protein content from 6.1% (without optimization) to 13.2%. We further applied the optimized medium and conditions in high cell density cultivation, and determined that the soluble target protein constituted 10.5% of the total protein. Our identification of the economic medium composition, optimal culture conditions, and details of the fermentation process should facilitate the potential application of ALF for further research. |
| 文献子类 | CNKI期刊论文 |
| 资助机构 | Supported by the National High Technology Research and Development Program of China(863 Program)(No.2006AA100311) |
| 卷 | v.29期:06页:105-115 |
| 语种 | 英文; |
| 分类号 | Q813 |
| ISSN号 | 0254-4059 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/191495]  |
| 专题 | 中国科学院海洋研究所 |
| 作者单位 | 1.R&DCenterofMarineBiotechnologyInstituteofOceanologyChineseAcademyofSciences 2.GraduateUniversityofChineseAcademyofSciences |
| 推荐引用方式 GB/T 7714 | Jiang S,Liu M,Wang BJ,et al. Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli. 2011. |
入库方式： OAI收割
来源：海洋研究所
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