Transcriptional analysis and regulation of carnobacteriocin production in Carnobacterium piscicola LV17

Abstract

Bacteriocin production by Carnobacterium piscicola LV17 (carnobacteriocin, Cbn) depends on the level of inoculation when grown in liquid medium. With an inoculum of ≥10 6 colony-forming units per ml (cfu/ml), bacteriocin production is observed during exponential growth, whereas with ≤10 4 cfu/ml no bacteriocin is detected even when the culture has reached stationary phase. Using pure bacteriocins, it was demonstrated that bacteriocin production is autoregulated. To understand how bacteriocin production is regulated at the molecular level, cell-free supernatant from a bacteriocin-producing culture was added to fresh medium at 1% (v/v) together with a non-producing inoculum (10 4 cfu/ml), to induce bacteriocin production (induced culture). Northern analysis revealed major transcripts of 0.35, 1.5 and 1 kb for carnobacteriocins A, B2 and BM1, respectively, indicating that regulation of bacteriocin production by inoculum size occurs at the transcriptional level. Primer extension demonstrated that transcription initiated from the same promoters with the induced culture as with the positive control (culture inoculated at 10 7 cfu/ml). Quantitative phosphorimager analysis of the primer extension products indicated that cbnA transcript was more abundant than cbnB2 or cbnBM1. © 1997 Elsevier Science B.V. All rights reserved.