title: Controlled intra- or extracellular production of staphylococcal nuclease and ovine omega interferon in Lactococcus lactis.
abstract: A system for controlled targeting of heterologous protein was developed in the food-grade bacterium Lactococcus lactis. It is composed of the L. lactis strain NZ9000 and of two broad host range expression vectors pCYT:Nuc and pSEC:Nuc for, respectively, cytoplasmic and secreted staphylococcal nuclease (Nuc) nisin-inducible production. The level of intracellular production of Nuc measured with pCYT:Nuc (3 mg x l(-1)) is significantly lower than the one obtained with pSEC:Nuc ( approximately 20 mg x l(-1)). The secretion efficiency (SE) of Nuc is estimated to be approximately 70%, corresponding to approximately 15 mg of secreted Nuc x l(-1). Furthermore, we established that Nuc production continued in L. lactis 10 h after a 1-h nisin-pulse induction. This system was then used for intra- and extracellular production of a protein of therapeutical interest in L. lactis, the ovine interferon-omega (IFN-omega). The SE and the quantity of secreted active IFN-omega were evaluated respectively to be approximately 70% and approximately 1 mg x l(-1) ( approximately two-fold higher than the cytoplasmic form).
authors.affiliation: Unité de Recherches Laitières et de Génétique Appliquée, INRA, Domaine de Vilvert, 78352 Cedex, Jouy en Josas, France.
authors.forename: Luis G
authors.initials: LG
authors.lastname: Bermúdez-Humarán
authors.forename: Philippe
authors.initials: P
authors.lastname: Langella
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Commissaire
authors.forename: Sébastien
authors.initials: S
authors.lastname: Gilbert
authors.forename: Yves
authors.initials: Y
authors.lastname: Le Loir
authors.forename: René
authors.initials: R
authors.lastname: L'Haridon
authors.forename: Gérard
authors.initials: G
authors.lastname: Corthier
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001709:Biotechnology / Q000379:methods*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D007372:Interferons / Q000235:genetics* / Q000378:metabolism; D013294:Lactococcus lactis / Q000235:genetics* / Q000378:metabolism; D008836:Micrococcal Nuclease / Q000235:genetics* / Q000378:metabolism; D009561:Nisin / Q000494:pharmacology; D010957:Plasmids; D012756:Sheep
chemical_list: D000900:Anti-Bacterial Agents; D009561:Nisin; D007372:Interferons; D008836:Micrococcal Nuclease