Statistical optimization of bacoside A biosynthesis in plant cell suspension cultures using response surface methodology

The in vitro cultures of Bacopa monnieri show poor production of the anti-Alzheimer's drug, bacoside A. Therefore, suitable bioprocess optimization strategy was developed for callus induction from leaf explants (30 days), followed by callus proliferation (15 days). Central Composite Design was implemented to analyze the effect of pH, photoperiod, naphthalene acetic acid (NAA), and benzylaminopurine (BAP) concentration for maximum biosynthesis of bacoside A using leaf explants as well as callus explants as the inoculum. Using the CCD responses, it was predicted that the best biomass concentration of 4.56 ± 0.53 g/l DW and bacoside A production of 14.04 ± 1.31 mg/g DW can be obtained using 5.4 pH, 18 h/6 h L/D photoperiod regime, and 1.2 mg/l BAP in combination with 0.2 mg/l NAA. The kinetic parameter values for maximum specific growth rate (0.16/day), saturation constant (7.35 g/l), inhibition constant (120 g/l), biomass yield (0.011 g/g), maintenance coefficient (0.02 g/g/day), and growth-associated (0.627 mg/g) and non-growth-associated (1.096 mg/g/day) bacoside A formation constants were determined experimentally in batch cultures using optimized conditions.