EVALUATION OF PROCESS VARIABLES FOR THE PRODUCTION AND PURIFICATION OF α-AMYLASE FROM BACILLUS LICHENIFORMIS MTCC-1483TARANJEET SINGH AND JAGDISH SINGHAbstract Present study reports the production of α-amylase from immobilized Bacillus licheniformis MTCC- 1483. Maximum production was reported when 2.0% (w/v) starch, 1.0% (w/v) peptone, 0.5% ammonium nitrate (w/v), 0.5% (w/v) trisodium citrate and 1.5% (w/v) calcium carbonate were used as media components at pH 6.5 and 37 °C, respectively. The innovative finding of this study was the production of a specific enzyme (alpha-amylase) under substrate stress conditions. Central composite design concerning alpha- amylase purification from egg white matrix was established using design expert. This is a first report on the purification of alpha-amylase by egg white matrix. Under optimized condition, 3.6-fold purification and 72.1% recovery yield was obtained. The purified enzyme showed a molecular weight of 54.0 kDa, that displayed maximum activity at pH 7.0 and 60 °C. Moreover, the Km and Vmax of purified enzyme were found to be 0.16 mg/mL and 7.1 IU/mg, respectively using starch as a substrate.
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