Systems-level understanding of how Propionibacterium acidipropionici respond to propionic acid stress at the...

In previous work, three evolved Propionibacterium acidipropionici mutants with higher tolerant capacity of propionic acid (PA) were obtained by genome shuffling. Here, we attempted to unravel the acid-tolerant mechanism of P. acidipropionici by comparing the physiological changes between P. acidipropionici and three mutants. The parameters used for comparison included intracellular pH (pHi), NAD⁺/NADH ratio, H⁺-ATPase activity, and the intracellular amino acids concentrations. It was indicated that the acid tolerance of P. acidipropionici was systematically regulated. Specifically, low pHi promoted the P. acidipropionici to biosynthesize more H⁺-ATPase to pump the protons out of the cells, and as a result, the NAD⁺/NADH ratio increased due to the decreased protons concentration. The increased arginine, aspartic acid, and glutamic acid concentrations helped to resist the acidic environment by consuming more H⁺ and generating more ATP and NH₃. Based on what was analyzed above, 20 mM arginine and aspartic acid were added during the shaker culture of P. acidipropionici, and the maximal PA titer reached 14.38 g/L, which was increased by 39.9% compared with the control.