1887

Abstract

The genome of ATCC 25745 contains a gene cluster that resembles a regulated bacteriocin system. The gene cluster has an operon-like structure consisting of a putative pediocin-like bacteriocin gene (termed ) and a potential immunity gene (termed ). Genetic determinants involved in bacteriocin transport and regulation are also found in proximity to and but the so-called accessory gene involved in transport and the inducer gene involved in regulation are missing. Consequently, this bacterium is a poor bacteriocin producer. To analyse the potency of the putative bacteriocin operon, the two genes were heterologously expressed in a host that contains the complete apparatus for gene activation, maturation and externalization of bacteriocins. It was demonstrated that the heterologous host expressing and produced a strong bacteriocin activity; in addition, the host became immune to its own bacteriocin, identifying the gene pair as a potent bacteriocin system. The novel pediocin-like bacteriocin, termed penocin A, has an isotopic mass [M+H] of 4684.6 Da as determined by mass spectrometry; this value corresponds well to the expected size of the mature 42 aa peptide containing a disulfide bridge. The bacteriocin is heat-stable but protease-sensitive and has a calculated pI of 9.45. Penocin A has a relatively broad inhibition spectrum, including pathogenic and species. Immediately upstream of the regulatory genes reside some features that resemble remnants of a disrupted inducer gene. This degenerate gene was restored and shown to encode a double-glycine leader-containing peptide. Furthermore, expression of the restored gene triggered high bacteriocin production in ATCC 25745, thus confirming its role as an inducer in the regulon.

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2006-06-01
2019-10-18
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Primers used in this study [PDF](10 kb) A comparison of inhibitory spectra of four different pediocin-like bacteriocins [PDF](25 kb)

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