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Study on the soluble exoression of recombinant human eoidermal growth factor using various fusion oartners in Escherichia coli  

Kim, Byung-Lip (Department of chemical Engineering, Chungnam National University)
Baek, Jung-Eun (Division of Biotechnology R&BD, Korea Research Institute of Bioscience and Biotechnology)
Kim, Chun-Sug (Division of Biotechnology R&BD, Korea Research Institute of Bioscience and Biotechnology)
Lee, Hyeok-Weon (Division of Biotechnology R&BD, Korea Research Institute of Bioscience and Biotechnology)
Ahn, Jung-Oh (Division of Biotechnology R&BD, Korea Research Institute of Bioscience and Biotechnology)
Lee, Hong-Weon (Division of Biotechnology R&BD, Korea Research Institute of Bioscience and Biotechnology)
Jung, Joon-Ki (Division of Biotechnology R&BD, Korea Research Institute of Bioscience and Biotechnology)
Lee, Eun-Gyo (Division of Biotechnology R&BD, Korea Research Institute of Bioscience and Biotechnology)
Kim, In-Ho (Department of chemical Engineering, Chungnam National University)
Publication Information
KSBB Journal / v.23, no.3, 2008 , pp. 205-212 More about this Journal
Abstract
The efficient soluble expression of human epidermal growth factor (hEGF) was achieved by using functional fusion partners in cytoplasm and periplasm of Escherichia coli (E. coli). hEGF was over-expressed in inactive inclusion body form in cytoplasm of E. coli due to improper disulfide bond formation and hydrophobic interaction, yielding about 5.9 mg/L in flask culture. Six functional fusion partners were introduced by linking to N-terminal part of hEGF gene for the high-level expression of soluble and active hEGF in cytoplasm and peri plasm region. Three fusion partners for cytoplasmic expression such as acidic tail of synuclein (ATS), thioredoxin (Trx) and lipase, and three fusion partners for periplasmic expression such as periplasmic cystein oxidoreductases (DsbA and DsbC) and maltose binding protein (MBP) were investigated. hEGF fused with ATS and DsbA showed over 90% of solubility in cytoplasm and periplasm, respectively. Especially DsbA was found to be an efficient fusion partner for soluble and high-level expression of hEGF, yielding about 18.1 mg/L and three-fold higher level compared to that of insoluble non-fusion hEGF in cytoplasm. Thus, heterologous proteins containing complex disulfide bond and many hydrophobic amino acids can effectively be produced as an active form in E. coli by introducing a suitable peptide or protein.
Keywords
hEGF; fusion partner; soluble expression; E. coli; cytoplasm; periplasm;
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