Heterologous expression of the Lactococcus lactis bacteriocin, nisin, in a dairy Enterococcus strain.
The bacteriocin nisin is produced only by some strains of Lactococcus lactis, and to date production in other lactic acid bacteria has not been achieved. Enterococcus sp. strain N12beta is a nisin-immune transconjugant obtained from a nisin-producing donor (L. lactis ATCC 11454) and a dairy recipient (Enterococcus sp. strain S12beta), but it does not produce nisin. In this study, using PCR amplification, we confirmed that the whole nisin operon is likely present in Enterococcus sp. strain N12beta. Northern hybridization of total RNA from strain N12beta with a nisA probe and the results of reverse transcriptase PCR showed the lack of nisA transcription in this strain. However, nisA transcription was partially restored in strain N12beta upon growth in the presence of exogenous nisin, and the nisA transcription signal was intensified after an increase in the external nisin level. Furthermore, bioassays showed that active nisin was produced in a dose-dependent fashion by strain N12beta following induction by exogenous nisin. These results indicated that expression of the nisin genes in Enterococcus sp. strain N12beta depended on autoinduction via signal transduction. However, the amount of external inducing signal required was significantly greater than the amount needed for autoinduction in L. lactis.
Li
Haiping
H
Department of Food Science and Nutrition, University of Minnesota, 1334 Eckles Avenue, St. Paul, MN 55108, USA.
O'Sullivan
Daniel J
DJ
D001430:Bacteriocins / Q000096:biosynthesis* / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D010450:Endopeptidases / Q000378:metabolism; D016983:Enterococcus / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D007109:Immunity; D007700:Kinetics; D013294:Lactococcus lactis / Q000235:genetics; D009561:Nisin / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis; D014158:Transcription, Genetic; D014169:Transformation, Bacterial
D001430:Bacteriocins; D011994:Recombinant Proteins; D009561:Nisin; D010450:Endopeptidases