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Secretion of biologically active murine interleukin-10 by Lactococcus lactis
Schotte1, Steidler, Vandekerckhove
1Department of Molecular Biology, Flanders Interuniversity Institute for Biotechnology, Ghent University, K.L. Ledeganckstraat 35, B-9000, GENT, Belgium
Enzyme and Microbial Technology
|December 19, 2000
Summary
Lactococcus lactis efficiently secretes biologically active murine interleukin-10 (mIL-10) as a fusion protein. Optimization of growth medium and pH increased mIL-10 yield by tenfold, achieving up to 30 mg per 10-litre fermentation.
Area of Science:
- Microbiology
- Biotechnology
- Immunology
Background:
- Interleukin-10 (IL-10) is a crucial immunosuppressive cytokine.
- Efficient secretion of therapeutic proteins by microbial hosts is a key goal in biotechnology.
Purpose of the Study:
- To investigate the secretion of biologically active murine interleukin-10 (mIL-10) by Lactococcus lactis.
- To optimize conditions for enhanced mIL-10 production.
Main Methods:
- Engineered mIL-10 as a fusion protein with the Usp45 secretion signal.
- Analyzed secreted protein using PAGE, ELISA, and bioassay.
- Optimized fermentation medium and pH for maximal yield.
Main Results:
- Lactococcus lactis efficiently secreted biologically active mIL-10.
- Confirmed correct processing of the fusion protein by lactococcal signal peptidase.
- Achieved a tenfold increase in mIL-10 yield by optimizing growth conditions, reaching up to 30 mg/L.
Conclusions:
- Lactococcus lactis is a viable host for the secretion of functional murine interleukin-10.
- Optimized fermentation strategies significantly enhance the yield of secreted recombinant proteins.