Simple method to identify bacteriocin induction peptides and to auto-induce bacteriocin production at low cell density.
The production of some bacteriocins by lactic acid bacteria is regulated by induction peptides (IPs) that are secreted by a dedicated secretion system. The IP gene cbaX, for carnobacteriocin A production by Carnobacterium piscicola LV17A, and a presumptive IP gene (orf6), associated with the genetic locus for enterocin B production in Enterococcus faecium BFE 900, were fused to the signal peptide of the bacteriocin divergicin A from Carnobacterium divergens LV13 to access the general secretory pathway. The culture supernatants of C. piscicola UAL26 and Lactococcus lactis MG1363 containing either of these constructs were used to induce bacteriocin production by Bac(-) cultures of C. piscicola LV17A or E. faecium CTC492. The cbaX fusion product induced bacteriocin production by Bac(-) C. piscicola LV17A, but the orf6 fusion product did not induce bacteriocin production by E. faecium CTC492. This represents a relatively simple method of confirming the role of presumptive IPs. The transformation of C. piscicola LV17A with the CbaX gene under expression of the P32 promoter from L. lactis resulted in constitutive production of bacteriocin by either the dedicated transport apparatus or the general secretory pathway.
Franz
C M
CM
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Canada.
Stiles
M E
ME
van Belkum
M J
MJ
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D001430:Bacteriocins / Q000096:biosynthesis* / Q000235:genetics; D016984:Enterococcus faecium / Q000235:genetics / Q000378:metabolism*; D007777:Lactobacillaceae / Q000235:genetics / Q000378:metabolism*; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010453:Peptide Synthases / Q000235:genetics; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; D001430:Bacteriocins; C087892:cbnBA protein, Carnobacterium piscicola; D010453:Peptide Synthases; C026269:2,3-dihydroxybenzoate - serine ligase