Optimization of compatible solutes for improving survival of freeze-dried Lactobacillus delbrueckii subsp. bulgaricus using Box-Behnken design

Lactobacillus delbrueckii subsp. bulgaricus is widely used in yogurt as a starter. The freeze-drying process may cause bacteria death. In the present work, the effect of three solutes (NaCl, sorbitol, and sodium glutamate) in MRS on viability of L.bulgaricus during freeze-drying was investigated. The optimal combination of adequate solutes was chosen by Box-Behnken Design. The survival rate and viable counts in freeze-dried powder, as well as the viable counts in broth, were used as responses. The results revealed that the optimum combination of solutes in MRS broth were 0.50% NaCl, 0.19% sorbitol, and 0.06% sodium glutamate. Under these optimal conditions, the survival rate was 53.2±0.14%, the viable counts in freeze-dried powder was 8.51±0.23×10¹⁰ CFU/g, and the viable counts in broth was 6.05±0.19 ×10⁸ CFU/mL, which were increased by 17.18%, 15.94%, and 17.31%, respectively, compared to the control. This research demonstrated the possibility of viability improvement of L.bulgaricus, which may provide a feasible reference for industrial development.