Lactobacillus reuteri, as a potential probiotic approved by Food Drug Administration and Chinese Ministry of Public Health, has been widely used in food and fermentation industries. Thus, it is of remarkable importance to develop the strategy for its high-cell-density (HCD) fermentation. In this study, the effects of batched culture, batch-fed culture and concentrated fed-batch (CFB) on the biomass of L. reuteri were compared. Then, the number of viable bacteria with or without aeration and with or without maintaining a constant pH were studied during CFB fermentation. The results showed that tangential perfusion and CFB can greatly prolong the logarithmic duration of the strain, and the number of viable bacteria was as high as 2.98×1015 CFU/mL. Compared with batched culture and batch-fed culture, the cell dry weight (CDW) in CFB fermentation increased by 5.3 times and 3.6 times, respectively. Under the conditions of ventilation (with a stable oxygen level of 15%), a constant pH level of 5.5 and a flow rate setting at 40 mL/min in CFB fermentation, the number of viable bacteria could reach up to 8.1×1015 CFU/mL, with a final CDW of 60.96 g/L which showed an improvement by 2.72 times and 1.07 times in comparison to the culturing manners of non-aeration and non-maintenance of constant pH, respectively. Together, these results suggest that HCD fermentation of L. reuteri can be achieved by a specific strategy of tangential perfusion and CFB, with considerable advantages in terms of obtaining live bacteria and biomass, laying the groundwork for a low-cost and time-saving large-scale production of L. reuteri.
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