1887

Abstract

Production and immunity of the two-component lantibiotic lacticin 3147 is encoded by the 60·2 kb lactococcal plasmid pMRC01. A 12·6 kb region of this plasmid, containing ten genes in two divergently arranged gene clusters, has been subcloned in subsp. MG1363 and has been shown to confer both lacticin 3147 production and immunity. Further subcloning revealed that the smaller of the two clusters () confers immunity. Although the and genes are homologous to ABC transporters which confer immunity to other lantibiotics, deletion analysis indicates that they do not play a role in the immunity exhibited by this subclone in subsp. MG1363. Also, a deletion in (which resembles a family of transcriptional repressors) had no effect on immunity. The remaining gene, , encodes a 116 amino acid protein with a predicted membrane location which bears no homology to other bacteriocin immunity proteins. Confirmation of its role in immunity was obtained when it was observed that disruption of resulted in a complete loss of immunity. When was cloned into the expression vector pMG36e, the resulting construct conferred levels of immunity comparable to pMRC01. This confirmed that under the control of a strong promoter, the gene product alone is sufficient to confer lacticin immunity. In addition, heterologous expression of was observed in OG1X. On cloning behind a nisin-inducible promoter, it was observed that the level of immunity was dependent on nisin concentration. Using this construct, the authors have demonstrated a potential role for as food-grade selectable marker. Thus, LtnI appears to represent a new class of lantibiotic immunity proteins.

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2000-01-01
2020-04-02
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