METHOD AND AGENT FOR THE ENZYMATIC DETERMINATION OF GLUCOSE

Glucose is determined by a sequence of reactions comprising converting glucose into glucose-6-phosphate by means of a phosphate donor and a transferase, converting the glucose-6-phosphate into 6-phosphogluconic acid by means of triphosphopyridine-nucleotide in the presence of glucose-6-phosphoric ac...

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Bibliographische Detailangaben
Hauptverfasser:	HANS ULRICH BERGMEYER, HANS MOELLERING
Format:	Patent
Sprache:	eng
Schlagworte:	BEER BIOCHEMISTRY CHEMISTRY COMPOSITIONS OR TEST PAPERS THEREFOR CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES ENZYMOLOGY INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES MEASURING MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS METALLURGY MICROBIOLOGY MUTATION OR GENETIC ENGINEERING PHYSICS PROCESSES OF PREPARING SUCH COMPOSITIONS SPIRITS TESTING VINEGAR WINE
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Beschreibung
Zusammenfassung:	Glucose is determined by a sequence of reactions comprising converting glucose into glucose-6-phosphate by means of a phosphate donor and a transferase, converting the glucose-6-phosphate into 6-phosphogluconic acid by means of triphosphopyridine-nucleotide in the presence of glucose-6-phosphoric acid dehydrogenase, reduced triphosphopyridine-nucleotide being formed at the same time, and then determining the latter spectrophotometrically: characterised in that phosphate donor is a compound of the general formula:- in which R is an organic radical, said radical not possessing a negative charge in the immediate neighbourhood of the phospate group, and wherein the transferase is acylphosphate: glucose p transferase. Specified phosphate donors are the following phosphates:-benzoyl, nicotoyl, acetyl, propionyl, N-benzoyl, glycoyl, carbonyl, glutaroyl, succinyl, and 3-phosphoglyceric acid-1-phosphate. The phosphate donor may be synthesized immediately before glucose determination by reacting acylate ions and an acylate-phosphokinase (e.g. acetyl phosphate from an acetate, adenosine triphosphate by means of acetate - kinase, and carbamyl phosphate from carbamic acid and ATP by means of carbamatephosphokinase.) Examples describe the analysis of various fruit juices and blood for glucose. Fructose, if present, does not interfere. In one Example, the acyl phosphate used is derived from a glycocollacetate buffer of pH 9.0, and magnesium chloride is also be present. The sample to be determined is mixed with all the reactant except the acyl phosphate-glucose transferase, and the latter is then added to start the reaction. Glucose is determined spectrophotometrically after some minutes. The invention also comprises a composition comprising the above defined phosphate donor, acyl phosphate: glucose transferase, glucose-6-phosphate-dehydrogenase and triphosphopyridine nucleotide: or a similar composition in which the phosphate donor is replaced by adenosine triphosphate, acylate ions and an acylate-phosphokinase.