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Production of recombinant human epidermal growth factor in Bacillus subtilis
Journal of the Taiwan Institute of Chemical Engineers ( IF 5.5 ) Pub Date : 2019-11-20 , DOI: 10.1016/j.jtice.2019.10.024
Heng-Hui Su , Jian-Chy Chen , Po-Ting Chen

Human epidermal growth factor (hEGF) is a crucial factor in wound healing, and therefore has wide applications in the pharmaceutical and cosmetics industries. To improve the production of hEGF protein for the industrial scale, we established an hEGF gene construct for recombinant secretion by Bacillus subtilis. Examination of the secretion abilities of various signal peptides (SPs) revealed six SPs that could effectively guide hEGF secretion from B. subtilis into the culture medium. The highest production level was obtained when the hEGF gene was fused with the xynD SP (lipo type). Two copies of the hEGF expression cassette had the same effect on improving production yield as isopropyl-β-d-1-thiogalactopyranoside (IPTG) induction, and the highest yield of hEGF can reach 360 ± 9.41 mg/L. Moreover, high-purity hEGF was obtained using a His-tagged purification system. Overall, these results demonstrate that B. subtilis is a suitable cell factory for hEGF production with improved yield compared to previous systems.



中文翻译:

枯草芽孢杆菌中重组人表皮生长因子的产生

人表皮生长因子(hEGF)是伤口愈合的关键因素,因此在制药和化妆品行业具有广泛的应用。为了提高工业规模的hEGF蛋白的产量,我们建立了hEGF基因构建体,用于枯草芽孢杆菌的重组分泌。各种信号肽(SPs)分泌能力的检查揭示了六个SPs,它们可以有效地指导hEGF从枯草芽孢杆菌的分泌到培养基中。当hEGF基因与xynD SP(脂质型)融合时,获得了最高的生产水平。hEGF表达盒的两个副本与异丙基-β- d具有相同的提高产量的作用-1-硫代半乳糖吡喃糖苷(IPTG)诱导,hEGF的最高产量可达到360±9.41 mg / L。此外,使用带有His标签的纯化系统获得了高纯度的hEGF。总体而言,这些结果表明枯草芽孢杆菌是适合hEGF生产的细胞工厂,与以前的系统相比,产量更高。

更新日期:2019-11-21
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