Use of bioreactor plant cell cultivation as a novel technique for mass production of bio-pesticide

摘要

Azadirachtin has been recognized as an efficient bio-pesticide worldwide. The present protocol for azadirachtin production is by solvent extraction of the seeds of Azadirachta indica which features low variable yield and contamination problems. Bioreactor plant cell cultivation could be an attractive alternative technology for commercial production of azadirachtin. Callus cultures were initiated from the seeds of Azadirachta indica variety having high azadirachtin content. Suspension culture was then developed in the shake flasks from the selected cell line under statistically optimized medium composition and environmental conditions. Batch kinetics of the cell suspension culture was established in the stirred tank reactor under optimized cultivation conditions. In order to facilitate the design of appropriate feeding strategies in fed-batch and continuous mode of cultivation a mathematical model was developed using the batch kinetics. This was eventually used to design a suitable fed-batch cultivation strategy. Model was also used to derive best off-line continuous cultivation strategy with cell retention for maximum biomass and azadirachtin production. The experimental verification of which resulted in a biomass accumulation of 95.8 g/L and highest azadirachtin production of 380 mg/L in the culture medium.