Isolation of cellulolytic activities from Tribolium castaneum (red flour beetle)

Cellulolytic enzymes have immense potential to convert cellulosic biomass into useful products. Tribolium castaneum crude proteins were isolated to screen the cellulolytic activities. The activity was established by substrate-agar plate assay and confirmed by endoglucanase assay. Cellulolytic activity was further purified and characterized using the different chromatographic techniques and electrophoresis. Gel filtration chromatography showed the presence of multiple forms of enzyme activities with different molecular weights. Stability of enzyme activity was investigated at different temperatures and pH. Optimum pH for was found 4.8 at 40oC determined as optimum temperature. Gradually decreasing Enzyme activity remained half at 60oC. Zymography and SDS-PAGE showed the presence of multiple forms of endoglucanase activities (Cel I and Cel II) with molecular weight of 55 kDa and 35 kDa.