The catabolite control protein A (CcpA) is a kind of multi-effect regulatory protein. In the study, the effect of the inactivation of CcpA and aerobic conditions on the growth, metabolic production, and stress tolerance to heat, oxidative, and cold stresses in Lactobacillus delbrueckii ssp. bulgaricus was investigated. Results showed that inactivation of CcpA distinctly hindered growth. Total lactic acid concentration was significantly lower in aerobiosis for both strains and was lower for the mutant strain than L. bulgaricus. Acetic acid production from the mutant strain was higher than L. bulgaricus in aerobiosis compared with anaerobiosis. Enzyme activities, lactate dehydrogenase (LDH), phosphate fructose kinase (PFK), pyruvate kinase (PK), and pyruvic dehydrogenase (PDH), were significantly lower in the mutant strain than L. bulgaricus. The diameters of inhibition zone were 13.59 ± 0.02 mm and 9.76 ± 0.02 mm for L. bulgaricus in anaerobiosis and aerobiosis, respectively; and 8.12 ± 0.02 mm and 7.38 ± 0.02 mm for the mutant in anaerobiosis and aerobiosis, respectively. For both strains, cells grown under aerobic environment possess more stress tolerance. This is the first study in which the CcpA-negative mutant of L. bulgaricus is constructed and the effect of aerobic growth on stress tolerance of L. bulgaricus is evaluated. Although aerobic cultivation does not significantly improve growth, it does improve stress tolerance.
Algal carcass is a low-value byproduct of algae after its conversion to biodiesel. Dried algal carcass is rich in protein, carbohydrate, and multiple amino acids, and it is typically well suited for growth and acid production of lactic acid bacteria. In this study, Lactobacillus delbrueckii ssp. bulgaricus ATCC 11842 was used to ferment different algal carcass media (ACM), including 2% ACM, 2% ACM with 1.9% glucose (ACM-G), and 2% ACM with 1.9% glucose and 2g/L amino acid mixture (ACM-GA). Concentrations of organic acids (lactic acid and acetic acid), acetyl-CoA, and ATP were analyzed by HPLC, and activities of lactate dehydrogenase (LDH), acetokinase (ACK), pyruvate kinase (PK), and phosphofructokinase (PFK) were determined by using a chemical approach. The growth of L. bulgaricus cells in ACM-GA was close to that in the control medium (de Man, Rogosa, and Sharpe). Lactic acid and acetic acid contents were greatly reduced when L. bulgaricus cells were grown in ACM compared with the control medium. Acetyl-CoA content varied with organic acid content and was increased in cells grown in different ACM compared with the control medium. The ATP content of L. bulgaricus cells in ACM was reduced compared with that of cells grown in the control medium. Activities of PFK and ACK of L. bulgaricus cells grown in ACM were higher and those of PK and LDH were lower compared with the control. Thus, ACM rich in nutrients may serve as an excellent substrate for growth by lactic acid bacteria, and addition of appropriate amounts of glucose and amino acids can improve growth and acid production.
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