Relationship between the physiology of Enterobacter agglomerans CNCM 1210 grown anaerobically on glycerol and the culture conditions

In a preliminary study, levels of activity of enzymes involved in anaerobic glycerol catabolism by Enterobacter agglomerans grown in batch cultures regulated in a pH range of 6.5-8.0 were monitored. That study showed that activities of key enzymes of the downstream metabolism of glycerol - glyceradehyde-3-phosphate dehydrogenase (GAP-DH), lactate dehydrogenase and pyruvate formate lyase - were strongly dependent on the culture pH. To investigate the influence of pH on the physiology of the strain, E. agglomerans was grown anaerobically in a continuous culture supplied with glycerol as the sole carbon source and regulated at pH 8. A complete biochemical analysis was performed and was compared with that previously described for the continuous culture regulated at pH 7. A limitation of the glycolytic flux at the level of GAP-DH was demonstrated at high dilution rate, resulting in an overflow metabolism through the 1,3-propanediol formation pathway. Increasing the specific rate of glycerol consumption also resulted in enhanced lactate production due to limitation by the pyruvate decarboxylation step. Finally, changing the culture pH significantly modified the enzymatic profile of E. agglomerans, and it enabled the stability of the culture to be increased by preventing the accumulation in the fermentation broth of 3-hydroxypropionaldehyde, an inhibitory metabolite, when the glycerol supply was suddenly increased.