Bestimmung von Saccharose,Glucose und Fructose mit trägergebundenen Enzymen

In serial chemical analysis the application of free enzymes is a rather uneconomical method because after each determination expensive enzyme reagents are discarded together with the test mixture, although as biocatalysts they are completely regenerated at the end of the reaction. In vivo, enzymes are often fixed to cell structures and more than up to a thousand times effective as biocatalysts. Hence the idea to apply enzymes fixed to a suitable carrier for repeated in vitro usage promises some success. Recently several groups of analysts succeeded in immobilizing enzymes of the protein metabolism for preparative purposes. We immobilized the enzymes saccharose, hexokinase, phosphohexose-isomerase and glucose-6-phosphate-dehydrogenase at CNBr activated agarose and by means of this affinity absorption-method determined the concentration of sucrose solutions in a closed recycling column system. The reduced form of nicotinamide-dinucleotide-phosphate being the measuring agent of the reaction is regenerated by means of glutathion reductase. The results of our investigation show, that the determination of the three carbohydrates sucrose, glucose and fructose by immobilized enzymes is by far superior to the common batch procedure regarding accuracy, velocity and costs.