High Cell Density Cultivation of Bifidobacterium longum Using a Calcium Carbonate-Alginate Beads System

A $CaCO_3$-alginate beads system was developed for high cell density cultivation of Bifidobacterium longum and the cost-effective media were also screened. In batch process with $CaCO_3$, beads, two strains of B. longum showed both the highest viable cells and optical density in TPY medium, resulting in maximum optical density and viable cell counts of 12.40, $2.22{\times}10^10$ cfu/ml for B. longum ATCC 15707 and 13.71, $3.93{\times}10^10$ cfu/ml for B. longum HLC 3742. Released size distribution, according to $CaCO_3$-alginate bead size preparation, was smaller than others. These results were also examined by observing their morphology. The skim milk-based medium was most adequate to cultivate B. longum as the cheapest medium, and $10\%$ skim milk supplemented with $2\%$ glucose and $1\%$ yeast extract was a suitable medium, supporting the growth to $5.57{\times}10^10$ cfu/ml for ATCC 15707 and $6.82{\times}10^9$ cfu/ml for HLC 3742. During the long-term storage at $4^{\circ}C\;and\;-20{\circ}C$, B. longum cultivated with $CaCO_3$ beads had the highest stability. Consequently, $CaCO_3$-alginate beads buffer was found to be useful not only to cultivate B. longum but also to preserve cultures.