Production of disulfide bond-rich peptides by fusion expression using small transmembrane proteins of Escherichia coli

被引:0
作者
Ziwei Chang
Ming Lu
Yunqi Ma
Dong-Geon Kwag
Seo-Hyun Kim
Ji-Min Park
Bo-Hye Nam
Young-Ok Kim
Cheul-Min An
Huayue Li
Jee H. Jung
Jang-Su Park
机构
[1] Pusan National University,Department of Chemistry and Chemistry Institute of Functional Materials
[2] Qingdao Institute of Bioenergy and Bioprocess Technology,Key Laboratory of Biofuels
[3] Chinese Academy of Sciences,Biotechnology Research Division, Aquaculture Industry Department
[4] National Fisheries Research and Development Institute,College of Pharmacy
[5] Pusan National University,undefined
来源
Amino Acids | 2015年 / 47卷
关键词
Disulfide bond; Carrier protein; Mass spectrometry;
D O I
暂无
中图分类号
学科分类号
摘要
Recombinant expression in Escherichia coli allows the simple, economical, and effective production of bioactive peptides. On the other hand, the production of native peptides, particularly those rich in disulfide bonds, is a major problem. Previous studies have reported that the use of carrier proteins for fusion expression can result in good peptide yields, but few are folded correctly. In this study, two transmembrane small proteins in E. coli, YoaJ and YkgR, which both orientate with their N-termini in cytoplasm and their C-termini in periplasm, were used for fusion expression. The recombinant production of two peptides, asteropsin A (ASPA) and β-defensin (BD), was induced in the periplasm of E. coli using a selected carrier protein. Both peptides were expressed at high levels, at yields of approximately 5–10 mg/L of culture. Mass spectrometry showed that the resulting peptide had the same molecular weight as their natural forms. After purification, single peaks were observed by reversed phase high-performance liquid chromatography (RP-HPLC), demonstrating the absence of isoforms. Furthermore, cytoplasmically expressed fusion proteins with a carrier at their C-termini did not contain disulfide bonds. This study provides new carrier proteins for fusion expression of disulfide bond-rich peptides in E. coli.
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页码:579 / 587
页数:8
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