Visualization of barley beta-glucan degrading isozymes after gel isoelectric focusing.
A simple method to study the polymorphism of barley (Hordeum vulgare L.) beta-(1-3,1-4)-glucanases (specific enzymes of barley beta-glucan hydrolysis) is described. Proteins of a crude extract of germinated barley kernels were separated in an immobilized pH gradient in two pH ranges (pH 3-10.5 and pH 4-7). beta-glucanases were visualized by contact printing with a polyacrylamide gel containing beta-glucan or lichenan. Patterns of beta-glucanases were revealed by staining with Congo Red with resultant clear zones on a stained background. Various conditions of germination, extraction and visualization were investigated. New isozyme bands could be detected and their nature and origin discussed.
D000180:Acrylic Resins; D002021:Buffers; D044644:Endo-1,3(4)-beta-Glucanase; D005782:Gels; D018525:Germination; D005936:Glucans / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000032:analysis*; D001467:Hordeum / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000032:analysis*; D010940:Plant Proteins / Q000032:analysis*; D047071:beta-Glucans
D000180:Acrylic Resins; D002021:Buffers; D005782:Gels; D005936:Glucans; D007527:Isoenzymes; D010940:Plant Proteins; D047071:beta-Glucans; C016680:polyacrylamide gels; C067858:beta-glucan, (1-3)(1-4)-; D006026:Glycoside Hydrolases; D044644:Endo-1,3(4)-beta-Glucanase