The quantitative effects of sugar concentration, nitrogen concentration, EDTA, temperature, pH and time of fermentation on ethanol production were optimized using a Box-Wilson central composite design (CCD) experiment. It was found that palmyra jaggery (sugar syrup from the palmyra palm) is a suitable substrate for the production of high concentrations of ethanol using Saccharomyces cerevisiae NCIM 3090 by submerged fermentation. A maximum ethanol concentration of 129.4 g/l was obtained after optimizing media components and conditions of fermentation. The optimum values were a temperature of 26.2°C, pH of 8.4, time of fermentation of 4.2 days with 398.5 g of substrate/l, 3.

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A rapid and efficient method the exploiting affinity of a-amylase for its substrate starch is described. a-amylase from Bacillus licheniformis was purified to homogeneity by ammonium sulphate precipitation and affinity chromatography with 230-fold purification. The a-amylase adsorption to various starches was examined in order to screen its ability for highest binding to starch. The a-amylase was bound to starch more tenaciously, hence various eluants like maltose, soluble starch and high salts could not elute the bound a-amylase. However, the bound a-amylase was instantly eluted using 2% (w/v) dextrin. The purified enzyme showed a single polypeptide on SDS-PAGE, with a molecular weight of 58 kD. Western blot analysis confirmed the specificity of antibody raised against purified a-amylase.

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