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High-level expression and characterization of bioactive human truncated variant of hepatocyte growth factor in Escherichia coli

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Abstract

Hepatocyte growth factor (HGF) is an effective anti-fibrotic factor because of its bioactivity in inhibiting fibrosis-related proteins in the development of hepatic fibrosis. However, high-level production of bioactive mature form HGF is difficult because of its complex structure. Here, we report a non-fusion protein expression system to obtain truncated variant of N-terminal hairpin and first kringle domains of HGF (tvNK1) in Escherichia coli to determine its anti-fibrotic effects on hepatic stellate cells (HSCs). Under the selected conditions of cultivation and isopropyl-β-D-1-thiogalactopyranoside induction, the expression level of tvNK1 accounted for approximately 65 % of the total cellular protein and 50 % of fusion protein in the supernatant of whole cell lysates. The recombinant protein could be purified in one step with Ni2+-affinity chromatograph. Finally, about 65 mg recombinant tvNK1 was obtained from 1 l fermentation culture with no <95 % purity. In vitro, the final purified tvNK1 was shown to inhibit the proliferation of HSCs and decrease the mRNA and protein expression levels of fibrosis-related COL1A1 and α-smooth muscle actin genes.

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Acknowledgments

This study was supported by grants from The National Natural Sciences Foundation of China (No. 81070329), Open Project Program of Heilongjiang Key Laboratory of Tissue Injury and Repair (No. zdsys2013-07) and Foundation of Mudanjiang Medical University (No. 2010-41, No. 2011-13).

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Authors and Affiliations

  1. Heilongjiang Key Laboratory of Tissue Damage and Repair, Mudanjiang Medical University, Mudanjiang, 157011, People’s Republic of China

    Xiaohua Wang, Haifeng Liu & Yanhui Chu

  2. Laboratory of Pathogenic Microbiology and Immunology, Mudanjiang Medical University, Mudanjiang, 157011, People’s Republic of China

    Xiaohua Wang, Yang Liu, Yuting Li, Jinqiu Gui & Yanhui Chu

  3. Hongqi Hospital of Mudanjiang Medical University, Mudanjiang, 157011, People’s Republic of China

    Zhongmin Zhang

Authors
  1. Xiaohua Wang
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  2. Haifeng Liu
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  3. Zhongmin Zhang
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  4. Yang Liu
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  5. Yuting Li
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  6. Jinqiu Gui
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  7. Yanhui Chu
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Corresponding author

Correspondence to Yanhui Chu.

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Xiaohua Wang and Haifeng Liu have contributed equally to this work.

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Wang, X., Liu, H., Zhang, Z. et al. High-level expression and characterization of bioactive human truncated variant of hepatocyte growth factor in Escherichia coli . World J Microbiol Biotechnol 30, 2851–2859 (2014). https://doi.org/10.1007/s11274-014-1711-3

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  • DOI: https://doi.org/10.1007/s11274-014-1711-3

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