Study of Cheese Associated Lactic Acid Bacteria Under Carbohydrate-Limited Conditions Using D-Stat Cultivation

The metabolic pathways alternative to glycolytic energy (ATP) during growth of starter and nonstarter lactic acid bacteria were studied simulating the depletion of carbohydrates during cheese ripening. D-stat cultivation strategy with the gradual decrease of galactose concentration in tryptone-arginine feeding medium was used. With the decrease of galactose feeding, the biomass yield calculated on carbohydrate consumption (Y_X/HEX) and acetate/lactate production ratio of all strains increased. We assume that ATP and biomass yields improved by directing the pyruvate flow from lactate to acetate and that metabolic energy could be obtained either by producing acetate from carbohydrates or from arginine metabolism in ADI-positive strains. Four LAB strains out of eight produced ornithine from arginine indicating active arginine-deiminase (ADI) pathway. These ADI-positive strains achieved 3-10 times higher Y_X/HEX than ADI-negative strains in tryptone-arginine medium. Lactobacillus plantarum also used serine as an energy source. Starters and NSLAB strains using the amino acids arginine and serine or limited amounts of carbohydrates therefore have the potential to influence flavor production in cheese more efficiently.     

Study of Cheese Associated Lactic Acid Bacteria Under Carbohydrate-Limited Conditions Using D-Stat Cultivation

The metabolic pathways alternative to glycolytic energy (ATP) during growth of starter and nonstarter lactic acid bacteria were studied simulating the depletion of carbohydrates during cheese ripening. D-stat cultivation strategy with the gradual decrease of galactose concentration in tryptone-arginine feeding medium was used. With the decrease of galactose feeding, the biomass yield calculated on carbohydrate consumption (Y_X/HEX) and acetate/lactate production ratio of all strains increased. We assume that ATP and biomass yields improved by directing the pyruvate flow from lactate to acetate and that metabolic energy could be obtained either by producing acetate from carbohydrates or from arginine metabolism in ADI-positive strains. Four LAB strains out of eight produced ornithine from arginine indicating active arginine-deiminase (ADI) pathway. These ADI-positive strains achieved 3–10 times higher Y_X/HEX than ADI-negative strains in tryptone-arginine medium. Lactobacillus plantarum also used serine as an energy source. Starters and NSLAB strains using the amino acids arginine and serine or limited amounts of carbohydrates therefore have the potential to influence flavor production in cheese more efficiently.     

Study of the toxic effect of short- and medium-chain monocarboxylic acids on the growth of Saccharomyces cerevisiae using the CO2-auxo-accelerostat fermentation system

The effect of aliphatic monocarboxylic acids (formic, acetic, propionic, valeric, octanoic and decanoic acids) on the growth and metabolic activity of Saccharomyces cerevisiae S288C was studied, using continuous cultivation method - CO(2)-auxo-accelerostat with smooth increase in the concentration of added monocarboxylic acids. Slow increase in the concentration of these acids resulted in the rapid decrease in the growth yield (Y(ATP)) and specific growth rate (micro), however, the specific ATP production rate (Q(ATP)) increased or stayed almost constant. On the other hand, Q(ATP) decreased if the concentration of formic, acetic or decanoic acids was increased rapidly. The toxic effect of aliphatic monocarboxylic acids on the growth of S. cerevisiae was characterized and quantified from the respective dose-effect curves as the IC(50) value (mM) using two different endpoints: a decrease of 50% in the specific growth rate (IC(50 micro)) and a decrease of 50% in the growth yield based on ATP production (IC(50YATP)). The concentrations of formic, acetic, propionic, valeric, octanoic and decanoic acids causing the 50% reduction in the specific growth rate (IC(50 micro)) were, respectively, 18.1, 47.1, 33.6, 2.3, 0.16 and 0.07 mM. The IC(50 micro) values were notably lower (up to 5-fold) in case of a more rapid increase in the concentration of acid in the medium. The results of the CO(2)-auxo-accelerostat experiments show that the toxic effect depends not only on the nature of the monocarboxylic acid (lipophilicity) but also on the rate at which its concentration changes in the growth environment.     

The effect of temperature and pH on the growth of lactic acid bacteria: a pH-auxostat study

The growth of Streptococcus salivarius subsp. thermophilus, Lactococcus lactis subsp. lactis, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus acidophilus and Lactobacillus paracasei was studied in a pH-auxostat. The computer-controlled smooth change of pH or temperature was used to study the effect of pH and temperature on the culture characteristics (specific growth rate mu growth yield Y(ATP) and specific lactate production rate Q(ATP)). The behaviour of mu and Q(ATP) with an increase of temperature suggested that ATP production capacity was an important factor in determining the maximum growth rate of lactic acid bacteria (LAB). The decrease of Y(ATP) with increase of temperature resulted in the decrease of specific growth rate while Q(ATP) remained constant or even increased. With the decrease of pH, decrease of both Y(ATP) and Q(ATP) was observed. S. thermophilus St20 having the highest maximum specific growth rate of 2.2 h(-1) at 44 degrees C was the most acid sensitive strain. The L. paracasei E1H3 having the lowest mu(max) was the most tolerant strain to pH change. The behaviour of mixed culture of L. bulgaricus and S. thermophilus in milk in a pH-auxostat with pH-decrease was in agreement with the pure culture experiments of the same species. The study showed that pH-auxostat with smooth controlled change of pH and temperature is an effective method for determination of technological characteristics and comparative physiological study of LAB.