Metabolic and Transcriptomic Adaptation of Lactococcus lactis subsp. lactis Biovar diacetylactis in Response to Autoacidification and Temperature Downshift in Skim Milk

ABSTRACT For the first time, a combined genome-wide transcriptome and metabolic analysis was performed with a dairy Lactococcus lactis subsp. lactis biovar diacetylactis strain under dynamic conditions similar to the conditions encountered during the cheese-making process. A culture was grown in skim milk in an anaerobic environment without pH regulation and with a controlled temperature downshift. Fermentation kinetics, as well as central metabolism enzyme activities, were determined throughout the culture. Based on the enzymatic analysis, a type of glycolytic control was postulated, which was shared by most of the enzymes during the growth phase; in particular, the phosphofructokinase and some enzymes of the phosphoglycerate pathway during the postacidification phase were implicated. These conclusions were reinforced by whole-genome transcriptomic data. First, limited enzyme activities relative to the carbon flux were measured for most of the glycolytic enzymes; second, transcripts and enzyme activities exhibited similar changes during the culture; and third, genes involved in alternative metabolic pathways derived from some glycolytic metabolites were induced just upstream of the postulated glycolytic bottlenecks, as a consequence of accumulation of these metabolites. Other transcriptional responses to autoacidification and a decrease in temperature were induced at the end of the growth phase and were partially maintained during the stationary phase. If specific responses to acid and cold stresses were identified, this exhaustive analysis also enabled induction of unexpected pathways to be shown.

Regulation of Gycolytic Metabolism in Asparagus under Low-oxygen, High-carbon-dioxide Atmospheres

Asparagus (Asparagus officinallis L. `Giant Jersey') was stored a in flow-through system at 0°C under levels of O2 ranging from 0.1 to 21 kPa in combination with three levels of CO2 (0, 10 and 20 kPa) for 21 d. The resulting changes in RQ and soluble sugars were monitored. The levels of sucrose were higher at 0 kPa of CO2 and at O2 levels >2 kPa; however, those levels were extremely reduced at combinations of high CO2 and low O2. Glucose levels were higher at 0 kPa CO2 when O2 concentrations levels were >1.5 kPa compared to CO2 at 10 and 20 kPa. Fructose levels were maintained higher with CO2 at 20 kPa for all levels of O2, showing lower levels as CO2 decreased. Glycolytic intermediates were evaluated to support the sugar accumulation data. Phosphorylated intermediate levels were altered in spears by CO2 and O2 treatments. Glycolytic control point enzymes were analyzed and may account for sugar accumulation and/or degradation induced by the atmospheric treatments.