Deciphering Clostridium metabolism and its responses to bioreactor mass transfer during syngas fermentation
This study used 13 C tracers and dynamic labeling to reveal metabolic features (nutrients requirements, pathway delineation and metabolite turnover rates) of Clostridium carboxidivorans P7, a model strain for industrial syngas fermentation, and its implication with bioreactor mass transfer. P7 shows poor activity for synthesizing amino acids (e.g., phenylalanine) and thus, needs rich medium for cell growth. The strain has multiple carbon fxation routes (Wood-Ljungdahl pathway, pyruvate:ferredoxin oxidoreductase reaction and anaplerotic pathways) and Re-citrate synthase (Ccar_06155) was a key enzyme in its tricarboxylic acid cycle (TCA) pathway. High fuxes were observed in P7’s Wood-Ljungdahl pathway, right branch of TCA cycle, pyruvate synthesis, and sugar phosphate pathways, but the cells anabolic pathways were strikingly slow. In bioreactor culture, when syngas fowrate increased from 1 to 10mL/min, P7 strain produced same amount of total extracellular products (acids and alcohols) but high fowrate favored alcohol accumulation. This observation was due to the mass transfer limitation infuencing energy metabolism (CO/H2 oxidation for cofactor generations) more prominently than carbon fxation. When syngas fowrate increased from 10 of 20mL/min, the alcohol productivity was not improved and the labeling rate (~0.03h -1 ) of key metabolite acetyl-CoA reached to P7 strain’s metabolism limitation regime.