Screening Production and Application of Alkaline Protease Enzyme Producing Bacillus Cereus from Fish
Author : Geetha G and Archana CU
Abstract :
Alkaline protease are one of the most important enzymes in the biological world. Microbial production of alkaline protease is getting more attention from researchers due to their unique properties and substantial activity. The present study was to isolate protease producing bacteria from contaminated sardine fish sample collected from local habitat of Ernakulam. The isolation was done by serial dilution and plating methods. All the isolates were screened for proteolytic activity on Skim milk agar plate at 37˚C for 48hrs. Protease activities were determined by the formation of clear zone around the colonies on SMA medium. Protease activity was compared using protease assay. The selected isolate was identified as B. cereus using standard identification parameters such as Gram staining followed by molecular analysis. The16srDNA of the selected organism showed 100% similarity in the BLAST search with B. cereus. The activity of enzyme in purified sample and supernatant was measured by tyrosine standard curve and it was found to be as 0.48 U/ml/min respectively. This result showed that B. cereus is a good producer of extra cellular protease and it can be used as biocleaner or in detergent manufacture.
Keywords :
Alkaline protease, B. cereus, skim milk agar, proteolysis, protease assay, tyrosine standard curve, detergent
