Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Production of Soluble Human Granulocyte Colony Stimulating Factor in E. coli by Molecular Chaperones

  • PARK SO-LIM (Department of Biotechnology & Bioengineering, Pukyong National University) ;
  • SHIN EUN-JUNG (Department of Biotechnology & Bioengineering/Department of Biomaterial Control, Dong-Eui University) ;
  • HONG SEUNG-PYO (BioLeaders Corp.,) ;
  • JEON SUNG-JONG (Department of Biotechnology & Bioengineering/Department of Biomaterial Control, Dong-Eui University) ;
  • NAM SOO-WAN (Department of Biotechnology & Bioengineering/Department of Biomaterial Control, Dong-Eui University)
  • Published : 2005.12.01
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Abstract

The effects of coexpression of GroEL/ES and DnaK/DnaJ/GrpE chaperones on the productivity of the soluble form of human granulocyte colony stimulating factor (hG-CSF) in E. coli were examined. Recombinant hG-CSF protein was coexpressed with DnaK/DnaJ/GrpE or GroEL/ES chaperones under the control of the araB or Pzt-1 promoter, respectively. The optimal concentration of L-arabinose for the expression of DnaK/DnaJ/GrpE was found to be 1 mg/ml. When L-arabinose was added at $OD_{600}$=0.2 (early-exponential phase), soluble hG-CSF production was greatly increased. In addition, it was observed that the DnaK/DnaJ/GrpE and GroEL/ES chaperones had no synergistic effects on preventing aggregation of hG-CSF protein. Consequently, by coexpression of the DnaK/DnaJ/GrpE chaperone, the signal intensity of the hG-CSF protein band in the soluble fraction of cell lysate was increased from $3.5\%\;to\;13.9\%$, and Western blot analysis also revealed about a 4-5-fold increase of production of soluble hG-CSF over the non-induction case of DnaK/DnaJ/GrpE.

Keywords

References

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