Single-cell level based approach to investigate acetate metabolism during batch industrial fermentation

Acetate is a product of Escherichia coli overflow metabolism when the bacteria are grown under aerobic conditions and glucose is present in excessive amount. It is an undesirable byproduct that affects growth, physiology, and performance of E. coli when used in industrial bioprocesses; its accumulation causes the decrease of productivity as it represents a waste of carbon source that would otherwise be converted to biomass and product. As the acetate accumulation problem is of the utmost importance in batch fermentation processes, various strategies have been developed to explain, understand and control the overflow metabolism phenomenon in E. coli. Even though acetate formation by E. coli have been studied for more than three decades, the literature published presents the results based on the average measurement of the whole population. The averaged data can mask the distribution of the activity on the sub-population level. We hypothesized that during the fermentation process, bacterial subpopulation exist, which exhibit different metabolic strategies towards the acetate. In this study, pure culture of Escherichia coli MG1655 was used to investigate in situ acetate metabolism at single-cell level during glucose fermentation. Batch fermentations were performed in order to examine consecutive stages of acetate metabolism during the fermentation process (production, co-consumption with glucose, consumption as single substrate). Uptake of glucose and acetate at single-cell level was observed and measured by quantitative microautoradiography.