Biochemical limitations of Bacillus thuringiensis based biopesticides production in a wheat bran culture medium

Abstract

Bacillus thuringiensis, a gram-positive sporulating bacteria found in the environment, produces, during its sporulation phase, crystals responsible for its insecticidal activity, constituted of an assembly of pore-forming δ-endotoxins. This has led to its use as a biopesticide, an eco-friendly alternative to harmful chemical pesticides. To minimize production cost, one endemic Bacillus thuringiensis sv. kurstaki (Btk) strain Lip, isolated from Lebanese soil, was cultivated in a wheat bran (WB) based medium (IPM-4-Citrus project EC n° 734921). With the aim of studying the biochemical limitations of Btk biopesticide production in a wheat bran based medium, the WB was sieved into different granulometries, heat treated, inoculated with Btk Lip at flask scale, then filtered and separated into an insoluble and a permeate fractions. Several biochemical analyses, ie. bio performances, starch, elemental composition, total nitrogen and ashes, were then conducted on both fractions before and after culture. On a morphological level, two populations were distinguished, the fine starch granules and the coarse lignocellulosic particles. The biochemical analyses showed that both the raw and sieved WB have a similar proteins content (0.115 g/gdm WB), water content (0.116 g/gdm WB) and elemental composition (carbon: 45%, oxygen: 37%, nitrogen: 3%, hydrogen: 6%, ashes: 5%). The starch content was 17%, 14% and 34% and the fermentable fraction was estimated to 32.1%, 36.1% and 51.1% respectively for classes 2, 3 and 4. Both the elemental composition and Kjeldahl analyses showed that the nitrogen is the limiting nutrient of the culture.