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Applied Microbiology and Biotechnology

, Volume 74, Issue 2, pp 316–323 | Cite as

Production of recombinant mink growth hormone in E. coli

  • Jolanta SereikaiteEmail author
  • Alina Statkute
  • Mindaugas Morkunas
  • Kostas Radzevicius
  • Vitaliano Borromeo
  • Camillo Secchi
  • Vladas-Algirdas Bumelis
Biotechnological Products and Process Engineering

Abstract

Escherichia coli cells expressing mink (Mustela vison) growth hormone were grown in a batch fermentation process. The expression level was estimated to be 27% of the total cellular protein after 3 h of induction with 1 mM isopropyl β-d-thiogalactoside (IPTG). If the expression of mink growth hormone (mGH) was induced with 0.2 mM IPTG, the concentration of target protein was slightly lower and was found to be 23% at the same time after induction. mGH expressed as inclusion bodies was solubilized in 8 M urea and renatured by dilution protocol at a protein concentration of 1.4–2.1 mg/ml in the presence of glutathione pair in a final concentration of 11.3 mM. [GSH]/[GSSG] ratio equal to 2/1 was used. Two-step purification process comprising of ion-exchange chromatography on Q-Sepharose and hydrophobic chromatography on Phenyl-Sepharose was developed. Some 25–30 mg of highly purified and biologically active mGH was obtained from 4 g of biomass. The method presented in this study allows producing large quantities of mGH and considering initiation of scientific investigation on mGH effect on mink in vivo and availability in fur industry.

Keywords

Growth Hormone Inclusion Body Nonreducing Condition Total Cellular Protein Inclusion Body Formation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Jolanta Sereikaite
    • 1
    Email author
  • Alina Statkute
    • 1
  • Mindaugas Morkunas
    • 1
  • Kostas Radzevicius
    • 1
  • Vitaliano Borromeo
    • 2
  • Camillo Secchi
    • 2
  • Vladas-Algirdas Bumelis
    • 1
  1. 1.Department of Chemistry and Bioengineering, Faculty of Fundamental SciencesVilnius Gediminas Technical UniversityVilniusLithuania
  2. 2.Department of Veterinary Pathology, Hygiene and Health, Biochemistry and Physiology UnitUniversity of MilanMilanItaly

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