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Application of response surfacemethodology (RSM) for protease production fromenterococcus hirae and using algae as substrate

ChandranMasi, E.Balaji, J.Vigneshwar,N.Parthasarathy

Protease is an enzyme that degrades protein, which has varied commercial applications in various fields. Protease enzyme is produced by using algae as substrate by various potential bacterial strains isolated fromdairy effluent. Chlorella sp. is an SCP and has high protein content therefore its potentiality to used as substrate were explored. Chlorella sp. was treated with different concentrations of humic acid 0.1%-1.0%, the protein content in algae was estimated and it is observed the protein content has been increased to its peak at 0.5% humic acid treated algae. Protein content isolated from 0.1% - 0.5% were used as substrate for the production of protease. The maximum protein content found in 0.5% treated algae (880 µg/ml). Enterococcus hirae was found to be more potent than Pseudomonas sp. in the production of protease at all concentration, especially at 0.5% humic acid treated algae (172µg/ml). Acinetobacter pitti was found to be least producing than the other species. Media components were optimized to enhance the Protease production by Enterococcus hirae with Response surface methodology; three major factors such asAlgal protein, Yeast extract and beef extractswere optimized. The optimizedmedia,which resulted in the maximum amount of protease (290 µg/ml), consisted of Algal protein (70ml/L), Glucose (10g/L) and beef extracts (10g/L).