Statistical media optimization for cell growth and azadirachtin production in Azadirachta indica (A. Juss) suspension cultures

Abstract

Azadirachtin is one of the most widely used biopesticide originating from a plant source. Its production from plant cell cultivation was viewed to overcome constraints associated with its regular supply from the seed kernels. In order to select the effective carbon and nitrogen source, different concentrations of carbon (sucrose/glucose) and nitrogen (NO3−/NH4+ ratio) were studied in A. indica suspension culture. Glucose turned out to be a better carbon source over sucrose yielding high biomass (6.32g/L) and azadirachtin (11.12mg/L) content. Nitrate alone as nitrogen source was favorable for both biomass and azadirachtin accumulation. Plackett–Burman design was adopted to select the most important nutrients influencing the growth and azadirachtin accumulation in suspension culture. After identifying effective nutrients, Central Composite Design (CCD) was used to develop mathematical model equations, study responses and establish the optimum concentrations of the key nutrients for higher growth and azadirachtin production. A maximum of 15.02g/L biomass and 2.98mg/g azadirachtin was produced using optimum nutrient concentrations representing 99 and 96% validity of the model prediction with respect to biomass and azadirachtin, respectively. This optimized media can be used for cultivation of A. indica cells in bioreactor for mass production of azadirachtin.