Advanced Journal of Microbiology Research

ISSN 2736-1756

Advanced Journal of Microbiology Research Vol. 2017

Available online at http://internationalscholarsjournals.org/journal/ajmr

© 2017 International Scholars Journals

Full Length Research Paper

Isolation, identification and characterization of multiple enzyme producing actinobacteria from sediment samples of Kodiyakarai coast, the Bay of Bengal

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

CAS in Marine Biology, Annamalai University, India.

Accepted 25 February, 2017

Abstract

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.

Key words: Amylase, cellulase, protease, actinobacteria, Kodiyakarai, Streptomycetes alboniger.