Determination of glycolytic intermediates in a flow injection system using immobilized enzymes.
Some glycolytic enzymes (lactate dehydrogenase, pyruvate kinase, enolase and phosphoglyceromutase) were immobilized on a polyacrylamide-type bead polymer containing carboxylic functional groups activated by water-soluble carbodiimide. The immobilized enzymes were used for the determination of pyruvic acid, phosphoenolpyruvic acid, 2-phosphoglyceric acid and 3-phosphoglyceric acid in a flow injection system. The immobilized lactate dehydrogenase column was repeatedly employed for the determination of pyruvic acid in clinical samples. The results of the flow injection method accorded well in accuracy, sensitivity and reproducibility with those of soluble enzyme analysis.
Simon
L M
LM
Department of Biochemistry, József Attila University, Szeged, Hungary.
Kotormán
M
M
Boross
L
L
Szajáni
B
B
D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005988:Glyceric Acids / Q000032:analysis*; D006019:Glycolysis; D006801:Humans; D008863:Microspheres; D010728:Phosphoenolpyruvate / Q000032:analysis*; D011773:Pyruvates / Q000032:analysis / Q000097:blood; D019289:Pyruvic Acid
D004800:Enzymes, Immobilized; D005988:Glyceric Acids; D011773:Pyruvates; C008885:2-phosphoglycerate; D010728:Phosphoenolpyruvate; C005156:3-phosphoglycerate; D019289:Pyruvic Acid