Project description:Lactococcus lactis NZ9000 and its parent MG1363 are the most commonly used lactic acid bacteria for expression and physiological studies. We noted unexpected but significant differences in the growth behaviors of both strains. We sequenced the entire genomes of the original NZ9000 and MG1363 strains using an ultradeep sequencing strategy. The analysis of the L. lactis NZ9000 genome yielded 79 differences, mostly point mutations, with the annotated genome sequence of L. lactis MG1363. Resequencing of the MG1363 strain revealed that 73 out of the 79 differences were due to errors in the published sequence. Comparative transcriptomic studies revealed several differences in the regulation of genes involved in sugar fermentation, which can be explained by two specific mutations in a region of the ptcC promoter with a key role in the regulation of cellobiose and glucose uptake.
Project description:pBL1 is a Lactococcus lactis theta-replicating 10.9-kbp plasmid that encodes the synthetic machinery of the bacteriocin Lcn972. In this work, the transcriptomes of exponentially growing L. lactis with and without pBL1 were compared. A discrete response was observed with a total of ten genes showing significantly changed expression. Up-regulation of the lactococcal oligopeptide uptake system (opp) was observed, likely linked to a higher nitrogen demand required for Lcn972 biosynthesis. Striking, celB coding for the membrane porter IIC of the cellobiose-PTS and the upstream gene llmg0186 were down-regulated. Growth profiles for L. lactis strains MG1363, MG1363/pBL1 and MG1363ΔcelB grown in CDM-cellobiose confirmed slower growth of pBL1 and ΔcelB while no differences were scored on glucose. The presence of pBL1 shifted the fermentation products towards a mixed acid profile and promoted substantial changes in intracellular pool sizes for glycolytic intermediates in cellobiose-growing cells as determined by HPLC and NMR. Overall, these data support the genetic evidence of a constriction in cellobiose uptake. Notably, several cell wall precursors accumulated, while other UDP-activated sugars pools were lower, which could reflect rerouting of precursors towards the production of structural or storage polysaccharides. Moreover, slow cellobiose-growing cells and those lacking celB were more tolerant to Lcn972 than cellobiose adapted cells. Thus, down-regulation of celB could help to build-up a response against the antimicrobial activity of Lcn972 enhancing self-immunity of the producer cells.
Project description:Lactococcus lactis NZ9000 and its parent MG1363 are the most commonly used lactic acid bacteria for expression and physiological studies. We noted unexpected but significant differences in the growth behaviors of both strains. We sequenced the entire genomes of the original NZ9000 and MG1363 strains using an ultradeep sequencing strategy. The analysis of the L. lactis NZ9000 genome yielded 79 differences, mostly point mutations, with the annotated genome sequence of L. lactis MG1363. Resequencing of the MG1363 strain revealed that 73 out of the 79 differences were due to errors in the published sequence. Comparative transcriptomic studies revealed several differences in the regulation of genes involved in sugar fermentation, which can be explained by two specific mutations in a region of the ptcC promoter with a key role in the regulation of cellobiose and glucose uptake. MG1363 versus NZ9000 in 2 different culture media
Project description:pBL1 is a Lactococcus lactis theta-replicating 10.9-kbp plasmid that encodes the synthetic machinery of the bacteriocin Lcn972. In this work, the transcriptomes of exponentially growing L. lactis with and without pBL1 were compared. A discrete response was observed with a total of ten genes showing significantly changed expression. Up-regulation of the lactococcal oligopeptide uptake system (opp) was observed, likely linked to a higher nitrogen demand required for Lcn972 biosynthesis. Striking, celB coding for the membrane porter IIC of the cellobiose-PTS and the upstream gene llmg0186 were down-regulated. Growth profiles for L. lactis strains MG1363, MG1363/pBL1 and MG1363ΔcelB grown in CDM-cellobiose confirmed slower growth of pBL1 and ΔcelB while no differences were scored on glucose. The presence of pBL1 shifted the fermentation products towards a mixed acid profile and promoted substantial changes in intracellular pool sizes for glycolytic intermediates in cellobiose-growing cells as determined by HPLC and NMR. Overall, these data support the genetic evidence of a constriction in cellobiose uptake. Notably, several cell wall precursors accumulated, while other UDP-activated sugars pools were lower, which could reflect rerouting of precursors towards the production of structural or storage polysaccharides. Moreover, slow cellobiose-growing cells and those lacking celB were more tolerant to Lcn972 than cellobiose adapted cells. Thus, down-regulation of celB could help to build-up a response against the antimicrobial activity of Lcn972 enhancing self-immunity of the producer cells. The transcriptomes of Lactococcus lactis MG1614 with and without the bacteriocinogenic plasmid pBL1, grown under laboratory conditions, were compared using three biological replicates.
Project description:Gene expression in Lactococcus lactis MG1363 was compared to that of L. lactis MG1363 â??guaA in rich GM17 medium. One condition design comparison of two strains