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Screening and selection of Streptomyces strains for efficient alkaline phosphatase (ALP) enzyme production

Pankaj Kumara, A.K. Roya, T.Kumara, M.Bhagata & B.N.Singhb
aPG. Department of Biotechnology, T. M. Bhagalpur University, Bhagalpur-812007,(Bihar)
bDept. of Botany, S. M. D degree College, J.P University, Chapra,(Bihar).
Received 11th January, 2011; Revised 28th February, 2011

Abstract : Alkaline phosphatase (APase; orthophosphoric-monoester phosphohydrolase,EC 3.1.3.1) is a hydrolase enzyme extremely important in molecular biology. It is present in most bacteria (Au et al., 1991)1, Archaea (Goldman et al., 1990)2, Yeast and Fungi (Furukawa et al., 1987)3. The different strains of Streptomyces bacteria were screened for alkaline phosphatase (ALP) enzyme production by resting cell assay method with the substrate paranitrophenylphosphate (PNPP) and by spectroscopic assay of yellow coloured byproduct, paranitrophenol is formed as a result of incubation of cell culture with the PNPP substrate. Out of the ten strains isolated from mangrove sediment; the S. sp. JS-12, S. venezuelae JS-11, S. bikiniensis JS-15 and S. sp. JS-20 were showed the good enzyme production when compared to the rest of the organisms. At the seventh day of incubation, the cultures were showed the yellow coloured byproduct formation due to the ALP enzyme action and the zone of diameter for the yellow coloured region contributes 34mm, 42mm, 50mm and 54mm in case of S. bikiniensis JS-15, S. venezuelae JS-11, S. sp. JS-12 and S. sp. JS-20 in the nitrate defined agar plate containing PNPP as a substrate with no external inorganic phosphate source. All these four strains exhibited the different level of production of ALP in the culture supernatant and in petriplates with medium containing no inorganic phosphate sources. The culture S. sp. JS-20 is the highest ALP enzyme producer when compared to other cultures. There was an increasing level of enzyme production as S. bikiniensis JS-15 < S. venezuelae JS-11 < S. sp. JS-2 < S. sp. JS-20.

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