Cereal Chem 53:157 - 173.  |  VIEW ARTICLE
Alpha-Amylase from Immature Hard Red Spring Wheat. I. Purification and Some Chemical and Physical Properties.

B. Marchylo, J. E. Kruger, and G. N. Irvine. Copyright 1976 by the American Association of Cereal Chemists, Inc. 

Alpha-Amylase was purified from immature Canadian hard red spring wheat by extraction, heat treatment at 70 C, acetone fractionation, complexing with glycogen, and DEAE-cellulose ion-exchange chromatography. Purity at the glycogen-complex stage was indicated by SDS gel electrophoresis. A total yield of 18.4% was obtained with 1900-fold purification. Three multiple forms of alpha-amylase were separated by DEAE- cellulose ion-exchange chromatography at pH 8.2. They were very similar in all of their properties, such as pH optimums, Km values, molecular weights, and activation energies, with the exception of their isoelectric points which were 4.65, 4.84, and 5.11 for the alpha-1, alpha-2, and alpha-3 components, respectively. The immature wheat alpha-amylases in having broader pH optimums, greater heat labilities, lower isoelectric points, and a higher molecular weight.