Overproduction of the Bacillus thuringiensis Vip3Aa16 toxin and study of its insecticidal activity against the carob moth Ectomyelois ceratoniae

Abstract The vip3Aa16 gene of Bacillus thuringiensis strain BUPM95 was cloned and expressed in Escherichia coli . Optimization of Vip3A16 protein expression was conducted using Plackett–Burman design and response surface methodology. Accordingly, the optimum Vip3A16 toxin production was 170 μg/ml at 18 h post-induction time and 39 °C post-induction temperature. This corresponds to an improvement of 21 times compared to the starting conditions. The insecticidal activity, evaluated against Ectomyelois ceratoniae , displayed an LC 50 value of 40 ng/cm 2 and the midgut histopathology of Vip3Aa16 fed larvae showed vacuolization of the cytoplasm, brush border membrane destruction, vesicle formati…