Thermostable alpha-amylase enzyme production from hot spring isolates Alcaligenes faecalis SSB17 – Statistical optimization

In the present investigation the production and characterization of a thermostable alpha-amylase enzyme from hot spring isolates Alcaligenes faecalis SSB17 has been studied. The individual and combined effect of temperature, pH and time has been investigated on the production of enzyme in four media (Luria Bertani, Nutrient Media, Semi-Synthetic Media l and Semi-Synthetic Media 2) using Box–Behnken design (BBD). Analysis of variance (ANOVA) shows high coefficient values of determination for each medium and satisfactory prediction second order regression models has been derived. Enzyme purification has been carried out by ammonium sulfate precipitation and size exclusion chromatography (SEC). Maximum purification has been obtained by SEC with 60 fold. SDS-PAGE of the purified enzyme has shown the presence of a single band associated with the amylase enzyme, having a molecular weight of 52 kDa