Enhanced production of Bacopa saponins by repeated batch strategy in bioreactor

Abstract

Cultivation of cell suspension culture of Bacopa monnieri targeting the production of bacosides was explored in a 5-l stirred tank reactor using statistically optimized conditions. The bioreactor cultivation conditions were modified and this led to profuse biomass growth (2.81 ± 0.20 g/l) and total bacosides (1.26 ± 0.23 mg/g in cells and 0.60 ± 0.11 mg/l in fermenter broth) production in 9 days. The values of static volumetric mass transfer coefficient (k_La), dimensionless mixing time (N_m) were measured in the bioreactor. The culture grew efficiently and produced enhanced amount of bacoside A (5.59 ± 0.41 mg/g total bacosides in cells and 3.12 ± 0.13 mg/l in the fermenter broth) using one cycle of repeated batch strategy adopted in the bioreactor for 15 days. The intracellular concentration of bacoside A3 (1.18 ± 0.11 mg/g), bacopaside II (2.09 ± 0.35 mg/g), bacopaside X (0.79 ± 0.17 mg/g) and bacopasaponin C (2.24 ± 0.23 mg/g) were significantly higher in repeated batch as compared to batch bioreactor cultivation. The yield of total bacosides in the fermenter broth was 5-times higher in repeated batch as compared to batch cultivation. This strategy can be helpful for the enhanced production of other valuable triterpenoid saponins.

Keywords: ANOVA; Bacoside A; Batch kinetics; Cell suspension culture; Repeated batch cultivation.