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Large-scale production of biologically active human keratinocyte growth factor-2

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Abstract

A rapid and efficient expression and purification system has been developed for large-scale production of biologically active recombinant human keratinocyte growth factor-2 (rhKGF-2). The gene encoding human KGF-2 was cloned into the expression vector pET3c and transformed into Escherichia coli BL21(DE3)/pLys S. Under optimal conditions in a 30-l fermentor, the average bacterial yield and the average expression level of rhKGF-2 of three batches were up to 732 g and 32%, respectively. The recombinant protein was purified by cation exchange and heparin-affinity chromatography. One hundred and sixty five milligrams of pure rhKGF-2 was achieved per liter culture. A preliminary biochemical characterization of purified rhKGF-2 was performed by Western blotting and mitogenic activity analysis, and the results demonstrated that purified rhKGF-2 could react with anti-human KGF-2 antibody and stimulate the proliferation of HaCat cells.

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Acknowledgments

The work was supported by grants from the National High-Tech Research and Development Program of China (2007AA02Z110) and the Science Foundation of Zhejiang Province of China (No. Z205755).

Author information

Correspondence to Xiaokun Li.

Additional information

Xiaoping Wu and Haishan Tian contributed equally to this work.

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Wu, X., Tian, H., Huang, Y. et al. Large-scale production of biologically active human keratinocyte growth factor-2. Appl Microbiol Biotechnol 82, 439–444 (2009). https://doi.org/10.1007/s00253-008-1782-x

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Keywords

  • Human keratinocyte growth factor-2
  • Large-scale production
  • Expression
  • Purification
  • Mitogenic activity