EXTRACELLULAR PROTEASE PRODUCTION BY ENTOMOPATHOGENIC FUNGUS Beauveria bassiana AND ITS COMPATIBILITY TEST AGAINST DETERGENT AND SURFACTANT

Beauveria bassiana is a well known entomopathogenic fungus which is considered as a non-selective bio-insecticide in agronomy. It acts as a parasite on a wide variety of arthropods. Protease is a group of different classes of enzyme that performs proteolysis; protein catabolism by hydrolysis of peptide bonds. Entomopathogenic fungi have the potentiality to produce extracellular protease as its combat mechanism against insect’s exoskeleton. Proteolysis activity was detected in the culture fluid when Beauveria bassiana was grown in a medium containing 2% casein as a sole carbon source, compared to that grown in Czapek Dox Broth (CDB) medium (sucrose less) supplemented with BSA, egg yolk, gelatin and maize extract in separate Erlenmeyer flasks. After protein extraction and purification, Sodium Dodecyl Sulfate (SDS) PAGE (12%) was used for 1-D electrophoresis analysis for the visualization of protein band by Coomassie Brilliant Blue staining. The amount of Protein was estimated by Bradford method before quantification of protease activity. The activity of protease was assessed by measuring the release of Trichloroacetic acid (TCA) soluble peptides from casein. From this research, maximum proteolytic activity was recorded in BSA mediated medium (2.00 U/ml) and the minimum activity was recorded in gelatin mediated medium (0.73 U/ml). Enzymatic activity of casein medium, egg yolk and maize extract supplemented casein medium were recorded as 1.25, 1.21 and 1.75 U/ml respectively. Protease from Beauveria strain retained nearly 87.5 - 97% in liquid shop and in case of hand wash that was 87.5 – 75%. Enzyme activity got lost when treated with laundry detergent and inhibitors. The best result was recorded in presence with surfactant, where 100% activity was retained. Although the application of such entomopathogenic fungi in agricultural field as the bio insecticide is established; but the finding of this research indicates one emerging application of the protease from B.bassiana as green detergent.