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A Comparative Analysis of Recombinant Protein Expression in Different Biofactories: Bacteria, Insect Cells and Plant Systems
Published on: March 23, 2015
Production processes of recombinant IL-1 beta from Bacillus subtilis: comparison between intracellular and
A V Bellini1, G Galli, E Fascetti
1Eniricerche S.p.A., Milan, Italy.
Journal of Biotechnology
|May 1, 1991
Summary
Intracellular expression of human interleukin-1 beta (IL-1 beta) in Bacillus subtilis yielded significantly higher production levels compared to extracellular secretion. This demonstrates B. subtilis
Area of Science:
- Biotechnology
- Molecular Biology
- Microbial Genetics
Background:
- Interleukin-1 beta (IL-1 beta) is a key inflammatory cytokine.
- Bacillus subtilis is a well-established host for recombinant protein production.
Purpose of the Study:
- To compare intracellular versus extracellular production of human IL-1 beta in Bacillus subtilis.
- To optimize growth conditions for enhanced IL-1 beta yield.
- To develop and compare purification strategies.
Main Methods:
- Insertion of human IL-1 beta coding sequence into intracellular (pSM261) and extracellular (pSM320) expression vectors.
- Transformation of Bacillus subtilis SMS118 with recombinant plasmids.
- Cultivation in 1-liter bioreactors under optimized conditions.
- Analysis of IL-1 beta levels in biomass and supernatant.
- Development and comparison of purification processes.
Main Results:
- Intracellular expression (pSM261) achieved 1-2.7 mg/g cells (up to 40 mg/L).
- Extracellular secretion (pSM320) yielded 0.27 mg/g cells (6.7 mg/L).
- Intracellular production was significantly more efficient than secretion.
Conclusions:
- Intracellular expression of IL-1 beta in B. subtilis is a highly efficient strategy.
- B. subtilis can be effectively used for recombinant protein production without relying on secretion.
- Optimization of intracellular expression offers superior yields for IL-1 beta production.
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