Kinetic assay for alpha-amylase

C - Chemistry – Metallurgy – 12 – Q

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150/3.2

C12Q 1/00 (2006.01) C12Q 1/25 (2006.01)

Patent

CA 1086198

ABSTRACT OF THE DISCLOSURE An inorganic phosphate assay comprising: (a) performing coupled reactions, at a pH of from about 6 to about 8, which comprise: (i) reacting maltose with an inorganic phosphate specimen in the presence of maltose phosphorylase to form glucose and .beta.-D-glucose-1-phosphate; (ii) reacting .beta.-D-glucose-1-phosphate in the presence of .beta.-D-phosphoglucomutase to form glucose-6-phosphate: and (iii) reacting glucose-6-phosphate in the presence of glucose-6-phosphate dehydrogenase and a co-enzyme selected from a group consisting of .beta.-nicotinamide-adenine dinucleotide, .beta.-nicotinamide-adenine dinucleotide phosphate and mixtures thereof to form the reduced form of said co-enzyme and 6- phosphogluconate; and (b) measuring the production of said reduced co-enzyme, wherein the inorganic phosphate being measured is the limiting component and wherein a non-phosphate buffer is used to control said pH.

337816

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