Studying Some Properties of Amylases Extracted from `d'Anjou' Pears

Abstract

Several characteristics of amylases involved in starch degradation were studied in extracts from immature (30 days before harvest) `d'Anjou' pears (Pyrus communis L.). Enzyme activity was not detected until after at least 60 minutes of incubation in frozen or lyophilized tissues. Activity increased significantly after 90 minutes and increased linearly after 2 to 12 hours of incubation. Activity was greater, however, in frozen than in lyophilized tissues. Three buffers (acetate, tris-HCl, and imidazole-HCl) were used at a range of pH levels (4.6-8.2) to ascertain the optimum assay system. Highest specific activity was recorded with acetate buffer at pH 5.6. The Km value in this system was 1.43 × 10-3g·ml-1. Specific activity increased as Ca concentration in the reaction mixture increased from 1 to 15 mm CaCl2 but did not change as Ca concentration increased from 15 to 25 mm CaCl2. The `d'Anjou' pear amylases were purified 5.7-fold using ammonium sulfate fractionation.