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Enhanced Production and Characterization of Commercially Important Thermostable Amylolytic Enzyme

Abstract

In the present study, different soil samples were analyzed to find a potent amylase producer. We found a natural soil isolate, named as Pseudomonas sp. B5 – a Gram negative short rod, which gave maximum amylase production (12.79 μmole/ml/min) in production medium containing 3% soluble starch when incubated for 24 h at 150 rpm. Further characterization of partially purified amylase proved it to be an enzyme having molecular weight of 55,370 KDa with 1.8 μmole Km and 0.019 U/min Vmax. Enzyme showed pH optima of 5 with activity in wide range of temperature ranging from 30 oC - 100 oC. This amylase was stable up to 35 minutes at 100 oC. Effect of several inhibitors and metal ions on amylase activity was also checked. EDTA proved to be detrimental for the amylase where as presence of metal ions namely Ca+2 and Fe+3 showed increase in the activity of the amylase.

Sarita Sharma, Rachana Bhatt

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