Molecular chaperone GroEL-GroES enhances the soluble expression of biologically active ovine growth hormone in the prokaryotic system

Protein Expr Purif. 2022 Aug:195-196:106097. doi: 10.1016/j.pep.2022.106097. Epub 2022 Apr 22.

Abstract

Growth hormone (GH) plays important roles in growth and development of mammalian animals and is valuable for many applications. This study aimed to express and purify biological active recombinant ovine growth hormone (roGH) through prokaryotic expression system. The roGH coding sequence was ligated into the prokaryotic expression vector and transformed into Escherichia coli (E. coli) for protein expression. Factors that influence the roGH expression were examined and the appropriate culture temperature (20 °C) and inducer (IPTG) concentration (25 μM) were determined. To enhance the soluble expression of the protein, co-expression with the molecular chaperone GroEL-GroES was utilized and eventually achieved a high yield of soluble roGH expressed in E. coli. Further, the fusion tag in expressed target protein could be efficiently removed through thrombin-specific cleavage. The expressed roGH was identified by Western blotting and the LC-MS spectrum confirmed its molecular weight of 22749.22 Da. Finally, the purified roGH had an expected biological activity when assayed in cell models in vitro and experimental mouse in vivo. In conclusion, the present study established an efficient and simple approach to produce recombinant GH, and facilitate relevant research and applications.

Keywords: GroEL-GroES; Ovine growth hormone; Prokaryotic expression; Trx fusion tag.

MeSH terms

  • Animals
  • Chaperonin 10
  • Chaperonin 60 / genetics
  • Chaperonin 60 / metabolism
  • Escherichia coli / metabolism
  • Escherichia coli Proteins* / metabolism
  • Growth Hormone* / genetics
  • Growth Hormone* / metabolism
  • Heat-Shock Proteins / metabolism
  • Mice
  • Molecular Chaperones / metabolism
  • Recombinant Proteins
  • Sheep

Substances

  • Chaperonin 10
  • Chaperonin 60
  • Escherichia coli Proteins
  • GroE protein, E coli
  • Heat-Shock Proteins
  • Molecular Chaperones
  • Recombinant Proteins
  • Growth Hormone