Fermentation optimization and kinetic model for high cell density culture of a probiotic microorganism: Lactobacillus rhamnosus LS-8.

Abstract

To develop a practical food-grade medium and optimal fermentation process for the cost-effective fermentation of Lactobacillus rhamnosus LS-8, both culture medium and conditions were optimized by combining single-factor experimental design, Plackett-Burman design and Box-Behnken design. The medium was simplified to five ingredients (g/L): whey powder (62.5), maltose syrup (50), corn steep liquor (55), NaCl (1) and lysine (0.05), and the optimal culture conditions were initial pH (6.28), constant fermentation pH (4.7), neutralizing agent (NaOH), aeration rate (0.2v/v/min) and stirrer speed (200rpm). After culturing in this optimized medium and conditions, the cell density of L. rhamnosus LS-8 was improved to 4.5 × 109CFU/mL, which was elevated about 9 times higher than that obtained in MRS medium. Moreover, cell growth and substrate consumption kinetic constants were determined by the logistic equation and Luedeking-Piret model, and the R2 values from the model equation were 0.9900 and 0.9971, respectively, indicating that these models were able to simulate the growth and substrate consumption of L. rhamnosus LS-8 accurately. In addition, a high-efficient production process of L. rhamnosus LS-8 was developed by repeated-batch operation, which was verified by five cycles of fermentation with good stability and repeatability. In conclusion, the efficiency of L. rhamnosus LS-8 fermentation was greatly improved as well as the reduction of the cost using the medium and process developed in the present study.