NADH Peroxidase for hydrogen peroxide determination
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United States of America Patent
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Jan 29, 1980
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N/A
app pub date -
Jun 29, 1977
filing date -
Jul 27, 1976
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Expired
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Abstract
A novel peroxidase for reduced nicotinamide-adenine-dinucleotide is provided which peroxidase oxidizes reduced nicotinamide-adenine-dinucleotide with hydrogen peroxide to give nicotinamide-adenine-dinucleotide and water and is characterized by a Michaelis constant K.sub.M to hydrogen peroxide of 2.8.times.10.sup.-5 M and to reduced nicotinamide-adenine-dinucleotide of 1.7.times.10.sup.-5 M, measured at 25.degree. C. in 0.2M tris buffer of pH 6.0, containing 0.1M potassium acetate. The peroxidase can be prepared by liberating the enzyme from Streptococcus faecalis ATCC 8043 by digestion or by treatment with a surface-active agent and isolating same from the enzyme solution obtained. Hydrogen peroxide is determined in a simple reaction or in a coupled reaction with a specific oxidase by contacting the said peroxidase with the H.sub.2 O.sub.2 producing reaction mixture at a pH of from 6.0 to 9.0 and measuring the change of extinction as a measure of H.sub.2 O.sub.2 and of the concentration of nicotinamide-adenine-dinucleotide initially present in said reaction mixture.
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