Journal of Biotechnology & Biomaterials
Open Access

Abstract

An extracellular amylase producing strain designated as SMVDU-1 was isolated from Bhairon Ghati of Trikuta Hills located in the state of J&K.The strain was identified as Bacillus firmus by 16S rDNA sequencing.The enzyme production was optimized using various Carbon sources and it was found that soluble starch and wheat bran increased the enzyme production up to 151% and 220% respectively as compared to Luria Bertani broth.The growth profile of the strain was studied at 35 o Cfor 72 hours and it was found that the enzyme production was best after 24 hours of growth. The amylase enzyme from the supernatant of SMVDU-1 was partially purified and characterized. The enzyme showed optimum activity at a temperature of 65ºC and pH 7.0. The temperature stability of the enzyme was found to be in the range of range of 45ºC -80ºC and the pH stability is in the range of 6.0 ? 8.0. Effect of metal ions like Ca 2+ , Mn 2+ , Mg 2+ , Cu 2+ , and Ag 2+ on enzyme activity was studied and it was found that Mn 2+ and Ca 2+ ions at a concentration of 2mM increased the enzyme activity considerably. Effect of inhibitors on enzyme activity was also studied and it was found that 50% of the enzyme activity was retained when SDS was used at a concentration of 2%. EDTA at a concentration of 1mM and 5mM was found to decrease the enzyme activity. From the above studies it may be concluded that the enzyme under investigation could be exploited for biotechnological applications because of its temperature optima and temperature stability and thermostable amylases have a great role to play in starch industry.

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