ABSTRACT

Chromatofocusing was used to resolve germinated wheat α-amylase isoenzymes. Optimum resolution and recovery of wheat α-amylase isoenzymes was achieved with a polybuffer™ 742 dilution factor of 10x, a buffer pH interval of 7.4 to 4.8, a column bed height of 45 cm, a flow rate of 0.5 mL/min, eluent buffer containing 10−4 M CaCl2 and a temperature of 4°C. A maximum of six peaks were resolved under these conditions. The first 3 peaks eluted were composed of GIII α-amylase isoenzymes with only a trace of GII isoenzymes. The remaining 3 peaks were composed of GI isoenzymes. The usefulness of chromatofocusing for separation of germinated wheat α-amylase isoenzymes was assessed.