Production of recombinant mink growth hormone in E. coli

Area of Science:

• Biotechnology
• Molecular Biology
• Protein Expression

Background:

• Mink growth hormone (mGH) is crucial for animal growth and development.
• Efficient production of recombinant mGH is essential for research and potential applications in the fur industry.

Purpose of the Study:

• To develop a scalable and efficient method for producing recombinant mink growth hormone (mGH) in Escherichia coli.
• To optimize expression conditions and purification strategies for high-yield production of active mGH.

Main Methods:

• Escherichia coli cells were engineered to express mink growth hormone (mGH).
• Batch fermentation was employed to culture the engineered E. coli.
• Recombinant mGH was induced using isopropyl β-D-thiogalactoside (IPTG) at varying concentrations (1 mM and 0.2 mM).
• Inclusion bodies containing mGH were solubilized in 8 M urea and renatured via a dilution protocol.
• A two-step purification process involving ion-exchange chromatography (Q-Sepharose) and hydrophobic interaction chromatography (Phenyl-Sepharose) was utilized.

Main Results:

• Expression levels of mGH reached 27% of total cellular protein with 1 mM IPTG induction after 3 hours.
• Slightly lower expression (23%) was observed with 0.2 mM IPTG.
• Purification yielded 25-30 mg of highly purified and biologically active mGH from 4 g of biomass.
• The renaturation protocol was effective at protein concentrations of 1.4-2.1 mg/ml using a glutathione redox buffer ([GSH]/[GSSG] ratio of 2/1).

Conclusions:

• A robust and scalable method for producing recombinant mink growth hormone (mGH) in E. coli has been established.
• The developed purification strategy effectively yields highly purified and biologically active mGH.
• This production method facilitates further in vivo studies on mGH effects in mink and supports its potential use in the fur industry.