Production and Mode of Action of Lactocin 27: Bacteriocin from a Homofermentative
            <i>Lactobacillus</i>

Upreti, G.C.; Hinsdill, Ronald D.

doi:10.1128/aac.7.2.139

Cited by 139 publications

(86 citation statements)

References 20 publications

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“…A narrow inhibitory spectrum, proteinaceous nature, and bactericidal mode of action are typical characteristics of antimicrobial agents classified as *bacteriocins (23). Very few lactobacifli have been confirmed to produce true bacteriocins within these criteria (3,4,6,25,26). Only a single bacteriocin has been identified and characterized within the L. helveticus species.…”

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Characterization and purification of helveticin J and evidence for a chromosomally determined bacteriocin produced by Lactobacillus helveticus 481

Joerger¹,

Klaenhammer²

1986

J Bacteriol

356

170

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Lactobacillus helveticus 481 produced an antimicrobial agent active against five closely related species. The sensitive indicators included L. helveticus 1846 and 1244, L. bulgaricus 1373 and 1489, and L. lactis 970. The antimicrobial compound was active at neutral pH under aerobic or anaerobic conditions, was sensitive to proteolytic enzymes and heat (30 min at 100°C), and demonstrated a bactericidal mode of action against sensitive indicators. These data confirmed that antimicrobial activity of L. helveticus 481 was mediated by a bacteriocin, designated helveticin J. Production of helveticin J was maximized in an anaerobic fermentor held lit a constant pH'of 5.5. Ultrafiltration experiments on cWuture supernatants containing the bacteriocin revealed that helveticin J was present as an aggregate with a molecular weight in excess of 300,000. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of helveticin J purified through Sephadex chromatography resolved a 37,000-dalton p,rotein band with bacteriocin activity. L. helveticus 481 was shown to harbor a single 8-megadalton plasmid (pMJ1008). Isolates cured of pMJ1008 were phenotypically identical to plasmid-bearing cells in fermentation patterns, helveticin J activity, and unmunity spectra. The data provided evidence for a chromosomal location of helveticin J and host immunity determinants.Lactobacillus helveticus and other lactobacilli have been shown to inhibit only closely related species (3,5,10,25). A narrow inhibitory spectrum, proteinaceous nature, and bactericidal mode of action are typical characteristics of antimicrobial agents classified as *bacteriocins (23). Very few lactobacifli have been confirmed to produce true bacteriocins within these criteria (3,4,6,25,26). Only a single bacteriocin has been identified and characterized within the L. helveticus species. Upreti and Hinsdill (25,26) confirmed the production of a bacteriogcin, designated lactocin LP27, by L. helveticus LP27. Lactocin LP27 exhibited a narrow activity spectrum by inhibiting strains of L. acidophilus and L. helveticus. A proteinaceous nature was demonstrated by inactivation of the bacteriocin with trypsin and pronase. In contrast to a bactericidal mode of action typical of bacteriocins, lactocin LP27 had a bacteriostatic effect on the indicator, L. helveticus LS18. Protein synthesis was terminated by lactocin LP27, but DNA and RNA synthesis and ATP levels were not affected.Bacteriocin production and resistance phenotypes offer excellent markers for study of genetic linkages and transfer systems. However, there is little information available on bacteriocins, genetic linkages, and gene transfer systems in Lactobacillus species. Gram-negative and gram-positive bacteria commonly harbor plasmid-borne genetic determinants of bacteriocin production and of host cell bacteriocin immunity (17,19,20,23,27). However, all attempts to locate plasmid determinants responsible for bacteriocin production and host immunity in lactobacilli have been unsuccessful. Difficulties in curing lactobaci...

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confidence: 99%

Characterization and purification of helveticin J and evidence for a chromosomally determined bacteriocin produced by Lactobacillus helveticus 481

Joerger¹,

Klaenhammer²

1986

J Bacteriol

356

170

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“…Bacteriocin genes are typically plasmid borne (43), but the genetic determinants encoding helveticin J appear to reside on the chromosome (16). Evidence for chromosomally located bacteriocin genes has also been reported for lactacin B produced by Lactobacillus acidophilus N2 (2) and lactocin 27 produced by L. helveticus LS18 (44).…”

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Cloning, expression, and nucleotide sequence of the Lactobacillus helveticus 481 gene encoding the bacteriocin helveticin J

1990

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Lactobacilus helveticus 481 produces a 37-kDa bacteriocin called helveticin J. Libraries of chromosomal DNA from L. helveticus were prepared in Agtll and probed for phage-producing fusion proteins that could react with polyclonal helveticin J antibody. Two recombinant phage, HJ1 and HJ4, containing homologous inserts of 350 and 600 bp, respectively, produced proteins that reacted with antibody. These two phage clones specifically hybridized to L. helveticus 481 total genomic DNA but not to DNA from strains that did not produce helveticin J or strains producing unrelated bacteriocins. HJ1 and H14 lysogens produced 0-galactosidase fusion proteins that shared similar epitopes with each other and helveticin J. The intact helveticin J gene (hlv) was isolated by screening a library of L. helveticus chromosomal DNA in XEMBL3 with the insert DNA from phage 1P4 as a probe. The DNA sequence of a contiguous 3,364-bp region was determined. Two complete open reading frames (ORF), designated ORF2 and ORF3, were identified within the sequenced fragment. The 3' end of another open reading frame, ORF1, was located upstream of ORF2. A noncoding region and a putative promoter were located between ORF1 and ORF2. ORF2 could encode an 11,808-Da protein. The L. helveticus DNA inserts of the HJ1 and HJ4 clones reside within ORF3, which begins 30 bp downstream from the termination codon of ORF2. ORF3 could encode a 37,511-Da protein. Downstream from ORF3, the 5' end of another ORF (ORF4) was found. A BglM fragment containing ORF2 and ORF3 was cloned into pGK12, and the recombinant plasmid, pTRK135, was transformed into LactobaciUus acidophilus via electroporation. Transformants carrying pTRK135 produced a bacteriocin that was heat labile and exhibited an activity spectrum that was the same as that of helveticin J.

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“…Barefoot and Klaenhammer (4) reported that 63% of the Lactobacillus acidophilus strains they examined produced bacteriocins. Bacteriocins produced by L. acidophilus 11088, L. acidophilus N2, Lactobacillus helveticus 481, and L. helveticus LP27 have been purified and shown to constitute a heterogeneous class of antimicrobial peptides (5,25,33,42). Although efforts have been made to characterize the proteinaceous nature of these bacteriocins, limited information is available about the genetic determinants of bacteriocins from lactobacilli or other LAB.…”

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confidence: 99%

Cloning, phenotypic expression, and DNA sequence of the gene for lactacin F, an antimicrobial peptide produced by Lactobacillus spp

1991

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Lactacin F is a heat-stable bacteriocin produced by Lactobacillus acidophilus 11088. A 63-mer oligonucleotide probe deduced from the N-terminal lactacin F amino acid sequence was used to clone the putative laf structural gene from plasmid DNA of a lactacin F-producing transconjugant, L. acidophilus T143. One clone, NCK360, harbored a recombinant plasmid, pTRK160, which contained a 2.2-kb EcoRI fragment of the size expected from hybridization experiments. An Escherichia coli-L. acidophilus shuttle vector was constructed, and a subclone (pTRK162) containing the 2.2-kb EcoRI fragment was introduced by electroporation into two lactacin F-negative strains, L. acidophilus 89 and 88-C. Lactobacillus transformants containing pTRK162 expressed lactacin F activity and immunity. Bacteriocin produced by the transformants exhibited an inhibitory spectrum and heat stability identical to those of the wild-type bacteriocin. An 873-bp region of the 2.2-kb fragment was sequenced by using a 20-mer degenerate lactacin F-specific primer to initiate sequencing from within the lactacin F structural gene. Analysis of the resulting sequence identified an open reading frame which could encode a protein of 75 amino acids. The 25 N-terminal amino acids for lactacin F were identified within the open reading frame along with an N-terminal extension, possibly a signal sequence. The lactacin F N-terminal sequence, through the remainder of the open reading frame (57 amino acids; 6.3 kDa), correlated extremely well with composition analyses of purified lactacin F which also predicted a size of 51 to 56 amino acid residues. Molecular characterization of lactacin F identified a small hydrophobic peptide that may be representative of a common bacteriocin class in lactic acid bacteria.

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Production and Mode of Action of Lactocin 27: Bacteriocin from a Homofermentative Lactobacillus

Cited by 139 publications

References 20 publications

Characterization and purification of helveticin J and evidence for a chromosomally determined bacteriocin produced by Lactobacillus helveticus 481

Characterization and purification of helveticin J and evidence for a chromosomally determined bacteriocin produced by Lactobacillus helveticus 481

Cloning, expression, and nucleotide sequence of the Lactobacillus helveticus 481 gene encoding the bacteriocin helveticin J

Cloning, phenotypic expression, and DNA sequence of the gene for lactacin F, an antimicrobial peptide produced by Lactobacillus spp

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