| Abstract: | ![]()
Immunoaffinity chromatography was used to purify the high pl α-amylase (α-amylase II) in a one step procedure after fractionation of the whole barley malt extract on Sephadex G25. The immunoglobulin G (IgG) fraction of an immune serum specific for the malt α-amylase II was immobilized on Ultrogel. A mild desorption procedure was used, combining distilled water elution with an interrupted elution. The purification was achieved within half a day including kernel extraction. The quality of the purification was assayed by SDS polyacrylamide gel electrophoresis, crossed immunoelectrophoresis and isoelectric focusing. For the second technique, an immune serum was used which was polyspecific for malt proteins including the high pl α-amylase (α-amylase II). The effect of this procedure on the specific activity of the enzyme and on its antigenicity was evaluated. The results underline the efficiency of the purification procedure and indicate that α-amylase II accounts for a few percent of the total soluble protein in malts. However, the α-amylase II fraction was not completely free from α-amylase I. The procedure resulted in a partial loss of the enzymatic activity but not of the antigenicity. |
