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A new human growth hormone production process using a recombinant Bacillus subtilis strain
E Franchi1, F Maisano, S A Testori
1Eniricerche S.p.A., Milan, Italy.
Journal of Biotechnology
|April 1, 1991
Summary
Researchers engineered hybrid plasmids for human growth hormone (hGH) precursor synthesis in Bacillus subtilis. Specific N-terminal sequences influenced precursor solubility and enabled purification, yielding authentic hGH.
Area of Science:
- Biotechnology
- Molecular Biology
- Protein Engineering
Background:
- The synthesis of recombinant human growth hormone (hGH) is crucial for therapeutic applications.
- Challenges exist in expressing and purifying biologically active hGH in microbial systems.
Purpose of the Study:
- To develop a novel expression system for producing authentic human growth hormone (hGH) in Bacillus subtilis.
- To investigate the impact of N-terminal modifications on hGH precursor solubility and purification.
Main Methods:
- Construction of hybrid plasmids encoding hGH precursors with varied N-terminal extensions in Bacillus subtilis.
- Analysis of precursor solubility based on N-terminal sequence characteristics (e.g., charged residues, homopolymeric tails).
- Purification of soluble precursors using charged residues and subsequent cleavage with Factor Xa protease to obtain mature hGH.
Main Results:
- Different N-terminal sequences conferred distinct solubility properties to hGH precursors.
- Shorter, charged N-terminal extensions (e.g., MRR-IEGR, MEELM-IEGR) enhanced precursor solubility compared to Met-hGH.
- Soluble precursors were efficiently purified and processed to yield authentic hGH.
Conclusions:
- Engineered N-terminal sequences can significantly improve the solubility and purification of recombinant hGH precursors.
- This strategy provides a viable method for the production and purification of authentic hGH in Bacillus subtilis.
- The developed protocol offers a promising approach for large-scale hGH manufacturing.