Amylase Production Through Agro-Waste

Increased production of ¿-amylase was observed with Bacillus amyloliquefaciens in the present study with wheat flour among the various agro-byproducts tested. The amylase from the bacterium was optimally active at a temperature of 37°C for 72h at pH 4±0.2 by using solid substrate (wheat flour) with the initial moisture of 80% and inoculated with Bacillus amyloliquefaciens of ¿2×106 cfu ml-1 of inoculum size. Supplementation with 0.04M KH2PO4, 1% maltose as a carbon source, tryptone and ammonium chloride as inorganic and organic nitrogen source in the substrate enhanced the enzyme yield considerably. The ¿-amylase from Bacillus amyloliquefaciens was partially purified by ammonium sulphate precipitation and ion exchange chromatography. Purification of amylase enzyme yielding a 2.05 fold purified enzyme with a specific activity of 11214.70 IU/mg. Characterization of purified enzyme revealed maximum stability on 65¿C temperature at pH 6±0.2. The enzyme was stable against Ca+2, Fe+3, Mn+2 metal ions however Cu+2 showed inhibitory effect. The characteristics of the enzyme qualify it as a good enzyme in biotechnological applications.