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

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• 作者：Ziwei Chang ; Ming Lu ; Yunqi Ma ; Dong-Geon Kwag ; Seo-Hyun Kim ; Ji-Min Park…
• 关键词：Disulfide bond ; Escherichia coli ; Carrier protein ; Mass spectrometry
• 刊名：Amino Acids
• 出版年：2015
• 出版时间：March 2015
• 年：2015
• 卷：47
• 期：3
• 页码：579-587
• 全文大小：3,490 KB
• 参考文献：1. Burrowes OJ, Diamond G, Lee TC (2005) Recombinant expression of pleurocidin cDNA using the / Pichia pastoris expression system. J Biomed Biotechnol 4:374-84. doi:10.1155/JBB.2005.374 CrossRef
  2. de Marco A (2009) Strategies for successful recombinant expression of disulfide bond-dependent proteins in / Escherichia coli. Microb Cell Fact 8:26. doi:10.1186/1475-2859-8-26 CrossRef
  3. de Marco A (2012) Recent contributions in the field of the recombinant expression of disulfide bonded proteins in bacteria. Microb Cell Fact 11:129. doi:10.1186/1475-2859-11-129 CrossRef
  4. Fontaine F, Fuchs RT, Storz G (2011) Membrane localization of small proteins in / Escherichia coli. J Biol Chem 286(37):32464-2474. doi:10.1074/jbc.M111.245696 CrossRef
  5. Haider SR, Reid HJ, Sharp BL (2010) Modification of tricine-SDS-PAGE for online and offline analysis of phosphoproteins by ICP-MS. Anal Bioanal Chem 397(2):655-64. doi:10.1007/s00216-010-3588-9 CrossRef
  6. Heras B, Kurz M, Shouldice SR, Martin JL (2007) The name’s bond…disulfide bond. Curr Opin Struct Biol 17:691-98 CrossRef
  7. Huber D, Boyd D, Xia Y, Olma MH, Gerstein M, Beckwith J (2005) Use of thioredoxin as a reporter to identify a subset of / Escherichia coli signal sequences that promote signal recognition particle-dependent translocation. J Bacteriol 187(9):2983-991. doi:10.1128/Jb.187.9.2983-2991.2005 CrossRef
  8. Ingham AB, Moore RJ (2007) Recombinant production of antimicrobial peptides in heterologous microbial systems. Biotechnol Appl Biochem 47(Pt 1):1-. doi:10.1042/BA20060207
  9. Ito K, Inaba K (2008) The disulfide bond formation (Dsb) system. Curr Opin Struct Biol 18:450-58 CrossRef
  10. Kurokawa Y, Yanagi H, Yura T (2000) Overexpression of protein disulfide isomerase DsbC stabilizes multiple-disulfide-bonded recombinant protein produced and transported to the periplasm in / Escherichia coli. Appl Environ Microbiol 66(9):3960-965 CrossRef
  11. Lavallie ER, Diblasio EA, Kovacic S, Grant KL, Schendel PF, Mccoy JM (1993) A thioredoxin gene fusion expression system that circumvents inclusion body formation in the / Escherichia coli cytoplasm. Biotechnol 11(2):187-93. doi:10.1038/Nbt0293-187 CrossRef
  12. Li Y (2009) Carrier proteins for fusion expression of antimicrobial peptides in / Escherichia coli. Biotechnol Appl Biochem 54(1):1-. doi:10.1042/BA20090087 CrossRef
  13. Li H, Bowling JJ, Fronczek FR, Hong J, Jabba SV, Murray TF, Ha NC, Hamann MT, Jung JH (2013) Asteropsin A: an unusual cystine-crosslinked peptide from porifera enhances neuronal Ca2+ influx. Biochim Biophys Acta 1830(3):2591-599. doi:10.1016/j.bbagen.2012.11.015 CrossRef
  14. Malakhov MP, Mattern MR, Malakhova OA, Drinker M, Weeks SD, Butt TR (2004) SUMO fusions and SUMO-specific protease for efficient expression and purification of proteins. J Struct Funct Genomics 5(1-):75-6. doi:10.1023/B:JSFG.0000029237.70316.52 CrossRef
  15. Malik A, Rudolph R, Sohlin
• 作者单位：Ziwei Chang (1)
  Ming Lu (2)
  Yunqi Ma (1)
  Dong-Geon Kwag (1)
  Seo-Hyun Kim (1)
  Ji-Min Park (1)
  Bo-Hye Nam (3)
  Young-Ok Kim (3)
  Cheul-Min An (3)
  Huayue Li (4)
  Jee H. Jung (4)
  Jang-Su Park (1)
  
  1. Department of Chemistry and Chemistry Institute of Functional Materials, Pusan National University, Busan, 609-735, Korea
  2. Key Laboratory of Biofuels, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, 266101, China
  3. Biotechnology Research Division, Aquaculture Industry Department, National Fisheries Research and Development Institute, 408-1 Sirang-ri, Gijang-eup, Gijang-gun, Busan, 619-902, Republic of Korea
  4. College of Pharmacy, Pusan National University, Busan, 609-735, Korea
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Biochemistry
  Analytical Chemistry
  Biochemical Engineering
  Life Sciences
  Proteomics
  Neurobiology
  
• 出版者：Springer Wien
• ISSN：1438-2199

文摘

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 5-0?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.