Screening, Characterization and Purification of Cellulase from Mango Mealybug Drosicha stebbingi Green (Homoptera: Margarodidae)
Screening, Characterization and Purification of Cellulase from Mango Mealybug Drosicha stebbingi Green (Homoptera: Margarodidae)
Mubasshir Sohail*, Qadeer Ahmed Soomro, Raza Muhammad, Muhammad Usman Asif and Imran Rauf
ABSTRACT
Arthropods have progressed with efficacious approaches to utilize plant cellulose for energy development and its study would be of a great importance in the field of bioenergy and integrated pest management. The study was aimed to evaluate cellulolytic activity which was detected in the gut of mango mealybug, (Drosicha stebbingi). Initially, cellulolytic activity was detected from crude proteins using substrate-agar plate assay and laterally confirmed by endoglucanase assay. Enzyme activity measured using the glucose standard curve was 725 U/ml after acetone precipitation. Thermal stability and optimum temperature were found to be 60°C for maximum activity. Likewise, enzyme was found more stable at 6.0 pH. Various mono and divalent cations were studied against the cellulolytic activity exhibited significant enhancing and inhibitory effect upon different concentrations. Effect of inhibitors (β-mercaptoethanol, EDTA) and various plant extracts on the enzyme activity were also tested. Cellulase was further purified and characterized through chromatographic techniques and electrophoresis. Gel filtration chromatography showed the presence of multiple forms of enzyme activities with different molecular weights. SDS-PAGE showed the presence of endoglucanase activities with molecular weight of 57 kDa.
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