A signal peptide secretion-dependent bacteriocin from Carnobacterium divergens.
Divergicin A is a strongly hydrophobic, narrow-spectrum, nonlantibiotic bacteriocin produced by Carnobacterium divergens LV13. This strain of C. divergens contains a 3.4-kb plasmid that mediates production of, and immunity to, the bacteriocin. N-terminal amino acid sequencing of the purified divergicin A was used to locate the structural gene (dvnA). The structural gene encodes a prepeptide of 75 amino acids consisting of a 29-amino-acid N-terminal extension and a mature peptide of 46 amino acids. Directly downstream of dvnA there is a second open reading frame that encodes the immunity protein for divergicin A. Divergicin A has a calculated molecular mass of 4,223.89 Da. The molecular mass determined by mass spectrometry is 4,223.9 Da, indicating that there is no posttranslational modification of the peptide. The N-terminal extension of divergicin A has an Ala-Ser-Ala (positions -3 to -1) cleavage site and acts as a signal peptide that accesses the general export system of the cell (such as the sec pathway in Escherichia coli). This is the first bacteriocin of lactic acid bacteria to be reported that does not have dedicated maturation and secretion genes. Production of divergicin A was observed in heterologous hosts containing only the two genes associated with divergicin A production and immunity. Fusing alkaline phosphatase behind the signal peptide for divergicin resulted in the secretion of this enzyme in the periplasmic space and supernatant of E. coli.
Worobo
R W
RW
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Canada.
Van Belkum
M J
MJ
Sailer
M
M
Roy
K L
KL
Vederas
J C
JC
Stiles
M E
ME
D000595:Amino Acid Sequence; D001419:Bacteria / Q000235:genetics*; D001430:Bacteriocins / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D005798:Genes, Bacterial; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010957:Plasmids; D021382:Protein Sorting Signals / Q000235:genetics*
D001430:Bacteriocins; D017931:DNA Primers; D021382:Protein Sorting Signals; C093812:dvnA protein, Carnobacterium divergens