Application of different downstream processing methods and their comparison for the large-scale preparation of Bacillus thuringiensis var. israelensis after fermentation for mosquito control

Abstract

Bacillus thuringiensis var. israelensis, a gram positive, spore-forming bacillus, produces parasporal crystal protein during sporulation, which is toxic in the mosquito larvae gut. An efficient downstream processing method for separating the spore crystal complex (SCC) from the fermented broth of B. thuringiensis var. israelensis is required to achieve maximum mosquitocidal activity. The different downstream processing methods, viz., tangential flow ultra-filtration, continuous centrifugation and acid precipitation were compared for their efficiency in separating SCC from broth obtained from a pilot-scale fermentor (100 l capacity). Among the three downstream processing methods, tangential flow ultra-filtration yielded the maximum amount of biomass (53.3 g/l), maximum number of spores (2.30 × 10 18 CFU/ml) and highest level of larvicidal activity (LC 50 28 nl/ml) against Aedes aegypti Bora-Bora strain followed by continuous centrifugation and acid precipitation methods.