Research Article
Mahesh M, Somashekhar R, Pr
Abstract
The 35 bacterial strains were isolated from water and soil sample from Karnataka (India) and screened for their ability to secrete extracellular Alkaline Phosphatase (ALP). Among them, a potent strain Proteus mirabilis resembling to Proteus mirabilis strain M54 was confirmed by 16S rRNA gene sequencing, was selected for the optimization of enzyme production. The optimum production was at pH 7-8 and at temperature 35°C for 28 hrs incubation. The pNPP was the best substrate for the ALP production. This production was stimulated by using starch and casein as sources of carbon and nitrogen respectively. After purification by Ammonium sulphate precipitation, gel filtration and ion exchange chromatography, the fold purification was increase up to 9.7. The molar mass was estimated by 12% SDS PAGE and was found to be approximately 56 kDa and also confirmed by gel filtration studies (data not shown). The study holds significance as there are only few reports available on the optimisation protocols.