Characterization of the nisin gene cluster nisABTCIPR of Lactococcus lactis. Requirement of expression of the nisA and nisI genes for development of immunity
TLDR
Transcription analyses of several L. lactis strains indicated that an expression product of the nisA gene, together with NisR, is required for the activation of nisinA transcription, indicating that NisI plays a role in the immunity mechanism.Abstract:
The nisin gene cluster nisABTCIPR of Lactococcus lactis, located on a 10-kbp DNA fragment of the nisin-sucrose transposon Tn5276, was characterized. This fragment was previously shown to direct nisin-A biosynthesis and to contain the nisP and nisR genes, encoding a nisin leader peptidase and a positive regulator, respectively [van der Meer, J. R., Polman, J., Beerthuyzen, M. M., Siezen, R. J., Kuipers, O. P. & de Vos, W. M. (1993) J. Bacteriol. 175, 2578–2588]. Further sequence analysis revealed the presence of four open-reading frames, nisB, nisT, nisC and nisI, downstream of the structural gene nisA. The nisT, nisC and nisI genes were subcloned and expressed individually in Escherichia coli, using the T7-RNA-polymerase system. This resulted in the production of radio-labelled proteins with sizes of 45 kDa (NisC) and 32 kDa (NisI). The nisT gene product was not detected, possibly because of protein instability. The deduced amino acid sequence of NisI contained a consensus Iipoprotein signal sequence, suggesting that this protein is a lipid-modified extracellular membrane-anchored protein. Expression of nisI in L. Iactis provided the cells with a significant level of protection against exogeneously added nisin, indicating that NisI plays a role in the immunity mechanism. In EDTA-treated E. coli cells, expression of nisI conferred up to a 170-fold increase in immunity against nisin A compared to controls. Moreover, a lactococcal strain deficient in nisin-A production, designated NZ9800, was created by gene replacement of nisA by a truncated nisA gene and was 10-fold less resistant to nisin A than the wild-type strain. A wild-type immunity level to nisin and production of nisin was obtained in strain NZ9800 harboring complementing nisA and nisZ plasmids. Transcription analyses of several L. IIactis strains indicated that an expression product of the nisA gene, together with NisR, is required for the activation of nisA transcription.read more
Citations
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Lantibiotic Nisin and Its Detection Methods
TL;DR: Two novel and highly sensitive nisin bioassays were developed based on NisRK mediated, nisin induced Green Fluorescent Protein (GFP) fl uorescence and the suitabilities of these assays for quantifi cation of nisin from food samples were evaluated in several food matrices.
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Intercellular Communication in Gram-Positive Bacteria Depends on Peptide Pheromones and Their Histidine Kinase Receptors
TL;DR: Interestingly, most peptide pheromone receptors belong to a newly defined subfamily of histidine kinases termed HPK 10 , indicating that they have evolved from a common ancestor.
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Lytr, a phage-derived amidase is most effective in induced lysis of Lactococcus lactis compared with other lactococcal amidases and glucosaminidases
Anton Steen,Saskia van Schalkwijk,Girbe Buist,Girbe Buist,Marja Twigt,Monika Szeliga,Wilco C. Meijer,Oscar P. Kuipers,Jan Kok,Jeroen Hugenholtz +9 more
TL;DR: Five genes encoding peptidoglycan hydrolases are present in the genome of Lactococcus lactis IL1403 and overexpression of lytR or the combination acmA/acmD resulted in increased cellular lysis in the cheese compared to the reference culture.
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Impact of the nisin modification machinery on the transport kinetics of NisT.
TL;DR: This study highlights the pivotal role of NisB for NisT in the secretion process in the presence and absence of the nisin maturation machinery, consisting of the dehydratase N isB and the cyclase NisC.
Functional analysis of the competence transcription factor ComK of Bacillus subtilis
TL;DR: In this paper, a ComK truncation variant lacking 9 aa from the N-terminal end (DeltaN9ComK) showed higher transcription activation than wild-type ComK when expressed in Lactococcus lactis.
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