| Summary: | Enzyme with relatively low stability shows an industrial incompetence which creates an urgent need of potential enzymes with
high stability in a cost effective way. Thus, thermostable enzymes derived from microorganism are the alternative resources for
convenient industrial application. Our present study was aimed for production of thermostable amylase with wide range of pH
and temperature stability, and induction of over production of the enzyme by using sugarcane bagasse as the sole carbon source.
The strain was an isolate from railway track of Cuttack railway station, Odisha, India. Morphologically the strain was identified
as Bacillus species by gram staining, and a potential amylase producer in normal Luria Bertani (LB) broth at 60°C. The optimum
extracellular amylase production observed was 0.085 U/ml after 6 hours at 60°C under submerged culture condition.
Extracellular amylase was precipitated through ammonium salt fractionation, and partially purified by Sephadex G-50 column
chromatography. The partially purified protein under SDS-PAGE was found to be resolved in to three distinct bands of molecular
weights approximately 108.69 kDa, 78.12 kDa and 65.63 kDa. The amylase enzyme showed wide range of temperature stability
till 90°C and pH stability from pH 5 to 8, respectively. However, the optimum stability of amylase was found at 90°C in pH 8.
The induction study has been carried out with abundantly available agro-industrial waste product sugarcane bagasse for over
production of amylase in four different concentrations (0.5%, 1.0%, 2.0%, and 5.0%). The enzyme production was enhanced with
increase in concentration of sugarcane bagasse as compared to the control LB medium. The highest amylase production observed
was 0.190 U/ml with 5.0% inducible substrate after 10 hours of submerged culture condition at 60°C. Thus, the strain is found to
be a potential producer of extracellular amylase and further studies are needed for the optimization of increased production and
induction of other possible enzymes by different inducible substrates.
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