Purification and characterization of a raw starch digesting alpha-amylase from the digestive juice of the snail limicolaria flammea (müller 1776)

An α-Amylase was purified to homogeneity from the digestive juice of L. flammea by Sephacryl S-200 HR gel filtration, DEAE-Sepharose CL-6B anion exchange and Phenyl-sepharose CL-6B hydrophobic interaction chromatography, with a 19.55-fold increase in specific activity and 7.8 % recovery. The molecular weight of the α-Amylase was estimated to be 62 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration. The optimum temperature and pH were 45°C and 5.0 respectively. The purified enzyme belonged to the EDTA-non sensitive α-amylase, but its activity was slightly stimulated by the presence of Ca2+ ions. The end-products of starch hydrolysis were glucose, maltohexaose and mor e oligosaccharides.