Open AccessControlled intra‐ or extracellular production of staphylococcal nuclease and ovine omega interferon in Lactococcus lactis
Author(s) -
BermúdezHumarán Luis G.,
Langella Philippe,
Commissaire Jacqueline,
Gilbert Sébastien,
Loir Yves,
L'Haridon René,
Corthier Gérard
Publication year - 2003
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(03)00475-0
Subject(s) - lactococcus lactis , extracellular , nisin , heterologous , microbiology and biotechnology , biology , secretion , nuclease , intracellular , cytoplasm , bacteria , biochemistry , lactic acid , enzyme , gene , genetics , antimicrobial
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 l −1 ) is significantly lower than the one obtained with pSEC:Nuc (∼20 mg l −1 ). The secretion efficiency (SE) of Nuc is estimated to be ∼70%, corresponding to ∼15 mg of secreted Nuc 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‐ω). The SE and the quantity of secreted active IFN‐ω were evaluated respectively to be ∼70% and ∼1 mg l −1 (∼two‐fold higher than the cytoplasmic form).