Improvement of the Lyophilization Survival Rate of Lactobacillus casei via Regulation of Its Surface Substances

Abstract

The influence of surface substance production on the freeze-drying survival of Lactobacillus casei and methods to control the surface substances during fermentation were studied. The bacteria were treated with hypertonicity combined with ultrasound, and the survival rate was determined. The optimal conditions for removing surface substance without harming the bacteria were 81 w/18 min. The surface substances provided a protective effect on the lyophilization of the bacteria without protectants. However, in the presence of protectants, excessive surface substances reduced the protective effect of the optimum protectant alginate to 39.69 ± 1.27%. Finally, the amount of surface substances and lyophilized survival rate of collected bacteria were determined by adding EDTA during fermentation and regulating fermentation conditions, such as the carbon source, carbon-to-nitrogen ratio, and pH. The highest survival rate was 85.79 ± 3.29%, which was achieved when the amount of surface substances was (2.82 ± 0.55) × 10−11 mg/CFU. Therefore, the production of surface substances by the bacteria could be reduced by modifying the fermentation stage, which has significance in the improvement of the lyophilization survival rate of L. casei and the number of live bacteria per unit mass of L. casei in the lyophilized preparation.