PURIFICATION AND CHARACTERIZATION OF THE COMMERCIALIZED, CLONED BACILLUS-MEGATERIUM ALPHA-AMYLASE .2. TRANSFERASE PROPERTIES

Using an assay procedure based on reduction of iodine binding to starch, Bacillus megaterium, alpha-amylase (BMA) demonstrated transferase activity using a wide range of acceptors. The enzyme had an absolute requirement for glucose or glucosides for acceptor molecules. Maltose acted as a transferase acceptor at low concentrations and as an inhibitor of starch hydrolysis at high concentrations. Kinetic analysis indicated that, in the presence of a suitable acceptor, the mechanism of starch hydrolysis is Ping Pong Bi Bi. The products of the transferase reaction have been determined using p-nitro-alpha-D-glucopyranoside as acceptor combined with a novel HPLC-based system for product detection.