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Genetic analysis of acidocin B : a novel bacteriocin produced by Lactobacillus acidophilus

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Author: Leer, R.J. · Vossen, J.M.B.M. van der · Giezen, M. van · Noort, J.M. van · Pouwels, P.H.
Type:article
Date:1995
Institution: Centraal Instituut voor Voedingsonderzoek TNO
Source:Microbiology, 7, 141, 1629-1635
Identifier: 37793
Keywords: Biology · Amino Acid Sequence · Bacteriocins · Base Sequence · Chromatography, High Pressure Liquid · Cloning, Molecular · Clostridium · Conserved Sequence · DNA, Bacterial · Genes, Bacterial · Lactobacillus · Lactobacillus acidophilus · Molecular Sequence Data · Multigene Family · Open Reading Frames · Plasmids · Protein Sorting Signals · Restriction Mapping · Support, Non-U.S. Gov't · Transformation, Bacterial

Abstract

The genes encoding the production of acidocin B, a bacteriocin produced by Lactobacillus acidophilus strain M46 which is active against Listeria monocytogenes, Clostridium sporogenes, Brochothrix thermosphacta, Lactobacillus fermentum and Lactobacillus delbrueckii subsp. bulgaricus, but inactive against most other Lactobacillus species, were previously localized on a 4 kb Xbal-HindIII fragment of plasmid pCV461. In the present work, DNA sequence analysis revealed the presence of three consecutive ORFs, which potentially code for hydrophobic peptides composed of 60, 91 and 114 amino acids, respectively, and a fourth ORF of opposite polarity which could potentially encode a peptide of 59 amino acids. The middle ORF (ORF-2; acdB) was identified as the gene encoding acidocin B by comparing the amino acid composition of highly purified acidocin B with the deduced amino acid sequence of ORF-2. Our results suggest that acidocin B is synthesized as a precursor which is processed at a site which conforms to the '-3, -1' rules of von Heijne to yield active acidocin B (59 amino acids). The presence of an immunity-protein-encoding gene on the 4 kb Xbal-BamHI fragment was deduced from the capacity of a plasmid vector harbouring this fragment to confer immunity upon transformation of L. fermentum NCK127. One of the three non-assigned ORFs may encode this immunity protein.