L. lactis P170 protein production system
L. lactis P170 production system
Bioneer's proprietary microbial protein production technology, the Lactococcus lactis P170 system, is used for production of high value recombinant proteins. The P170 system is available for licensing.
Among the main advantages of the P170 system which is based on the lactate-inducible promoter P170 is that the protein of interest is secreted to the culture medium simplifying the purification process. In turn, the production volume can be readily scaled up from a few mL to thousands of litres.
The P170 system includes expression vectors as well as production hosts based upon optimised L. lactis bacterial strains tailored to the P170 technology and fermentation procedures including chemically defined production media.
Bioneer enables clients to have their specific protein tested in the P170 system in order to assess if production in L. lactis is a feasible strategy. Such feasibility studies typically comprise:
• Construction of the production plasmid by either cloning the gene of interest directly from the source organism (cDNA or DNA) or as a codon-optimised gene into the desired P170 expression vector selected among a number of variants: low to high copy number variants, variants allowing the target protein to be secreted or to be expressed intracellularly, variants with purification tags, and variants with different selection markers.
• A number of L. lactis strains developed for protein production are transformed and tested for expression of the recombinant protein of interest. The initial expression studies take place in shaker flasks and/or 1 L fermentors, and the expression of the recombinant protein is analysed by standard protein analyses.
• As part of the documentation of the feasibility study, plasmid stability tests are performed on the production strain and a research Master Cell Bank is created.
Scientific publications on P170
- Flavobacterium sp. strain 4221 and Pedobacter sp. strain 4236 beta-1,3-glucanases that are active at low temperatures (Rasmussen, 2008) (PubMed)
- Production of Recombinant Peanut Allergen Ara h 2 using Lactococcus lactis (Glenting, 2007) (PubMed)
- Extracellular secretion of a maltogenic amylase from Lactobacillus gasseri ATCC33323 in Lactococcus lactis MG1363 and its application on the production of branched maltooligosaccharides (Cho, 2007) (PubMed)
- Safety of ESAT-6 (Aggerbeck, 2006) (PubMed)
- Two acid-inducible promoters from Lactococcus lactis require the cis-acting ACiD-box and the transcription regulator RcfB (Madsen, 2005) (PubMed)
- A Plasmodium falciparum GLURP-MSP3 chimeric protein; expression in Lactococcus lactis, immunogenicity and induction of biologically active antibodies (Theisen, 2004) (PubMed)
- Optimization of signal peptide SP310 for heterologous protein production in Lactococcus lactis (Ravn, 2003) (PubMed)
- The development of TnNuc and its use for the isolation of novel secretion signals in Lactococcus lactis (Ravn, 2000) (PubMed)
- Molecular characterization of the pH-inducible and growth phase-dependent promoter P170 of Lactococcus lactis (Madsen, 1999) (PubMed)¨
- Cloning and partial characterization of regulated promoters from Lactococcus lactis Tn917-lacZ integrants with the new promoter probe vector, pAK80 (Israelsen, 1995) (PubMed)
For further information, please contact Group Leader, Bacterial Expression; Søren Madsen by phone (+45 45160444) or email.