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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.
*Corresponding author.
E-mail:
manimaribtech@gmail.com.
Accepted 25 February, 2010 |
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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. |
