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
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Genetics of subtilin and nisin biosyntheses: biosynthesis of lantibiotics.
K D Entian,W M de Vos +1 more
TL;DR: Subtilin and nisin belong to the ribosomally synthesized peptide antibiotics, which are derived from prepeptides which are post-translationally modiffied and have been termed lantibiotics because of their characteristic lanthionine bridges.
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Gene Expression of Commensal Lactobacillus johnsonii Strain NCC533 during In Vitro Growth and in the Murine Gut
Emmanuel Denou,Emmanuel Denou,Bernard Berger,Caroline Barretto,Jean-Michel Panoff,Fabrizio Arigoni,Harald Brüssow +6 more
TL;DR: Major differences between in vitro- and in vivo-expressed genes in a Lactobacillus gut commensal organism are revealed but also marked changes in the expression of genes along the digestive tract.
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Restrictive Use of Detergents in the Functional Reconstitution of the Secondary Multidrug Transporter LmrP
TL;DR: Reconstituted LmrP mediated the transport of multiple drugs in response to an artificially imposed pH gradient, demonstrating that the protein functions as a proton motive force-dependent multidrug transporter, independent of accessory proteins.
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Potentiating the Activity of Nisin against Escherichia coli
TL;DR: This research presents a new and promising method to increase the anti-Gram-negative activity of lantibiotics and showed that when an eight amino acids (PRPPHPRL) tail from apidaecin 1b was attached to nisin, the activity of nisin against Escherichia coli CECT101 was increased more than two times.
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Evidence for a role of NisT in transport of the lantibiotic nisin produced by Lactococcus lactis N8
Mingqiang Qiao,Per E. J. Saris +1 more
TL;DR: Results demonstrate that NisT is the transport protein dedicated to translocate nisin and that dehydration and lanthionine formation in nisin maturation can occur independently of transport.
References
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