A new expression system for Lactococcus lactis was developed. The system is based on a phosphate starvation inducible pstF promoter of L. lactis MG1363. Intracellular β-galactosidase and secreted α-amylase were produced using this tightly regulated system. No evidence of regulatory sites in regions of the 5′-end of the pstF coding sequence was found. High expression levels of the β-galactosidase gene were obtained using the original pstF RBS in a phosphate-depleted medium. The results suggested that with the phosphate starvation inducible system, it is possible to achieve expression levels comparable to the ones obtained with the widely used nisin-controlled gene expression system (NICE). A specific β-galactosidase activity of 670 μkat g−1 using a phosphate-depleted medium and an α-amylase activity of 3.6 μkat l−1 in a bioreactor cultivation were produced. The advantages of the current expression system include that no prior removal of phosphate from the medium in bioreactor scale is required, and no additions of inducing agents are needed. Furthermore, the system can be operated in L. lactis without introduction of regulatory genes into the host.
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We thank Valio Ltd. and Soile Tynkkynen for cooperation and for the plasmid containing the lacZ gene.
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Sirén, N., Salonen, K., Leisola, M. et al. A new and efficient phosphate starvation inducible expression system for Lactococcus lactis . Appl Microbiol Biotechnol 79, 803–810 (2008). https://doi.org/10.1007/s00253-008-1484-4
- Lactococcus lactis
- Gene expression
- Phosphate starvation