Abstract

P. Manivasagan*, S. Gnanam, K. Sivakumar, T. Thangaradjou, S. Vijayalakshmi and T. Balasubramanian

A total of twenty nine strains were isolated from the Kodiyakarai coastal sediments (Bay of Bengal) and ten of them exhibited multiple enzyme activity. Of these, the best (GK-22) was selected based on zone formation (amylase, cellulase and protease) and its growth conditions were standardized for better production of multiple enzymes. Present study on production of multiple enzymes (amylase, cellulase and protease) by GK-22 showed higher enzyme levels at the end of the stationary phase after incubation for 72 h at pH 7.0. Maximum activity of amylase, cellulase and protease (84, 88 and 89 IU/ml, respectively) was obtained at pH 7.0, temperature 45°C, sodium chloride concentration 2%, carbon compound sucrose, nitrogen compound beef extract, amino acid L-asparagine for amylase and cellulase and L-tyrosine for protease. The multiple enzymes were purified by precipitation with ammonium sulphate and ion exchange chromatography and the SDS-PAGE showed a single band for the purified enzyme, with an apparent molecular weight of 80 (amylase), 66 (cellulase) and 97 KDa (protease). The strain, GK-22 which showed higher multiple enzyme activity was tentatively identified as Streptomyces alboniger. These findings suggest that the strain can effectively be used in large scale production of multiple enzymes for commercial purposes, after testing and ascertaining the strain’s capability in large scale fermentations.