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    • 10. 发明授权
    • NADH Peroxidase for hydrogen peroxide determination
    • NADH过氧化物酶测定过氧化氢
    • US4186052A
    • 1980-01-29
    • US811422
    • 1977-06-29
    • Albert RoderHans MolleringWolfgang GruberKlaus BeaucampHans SeidelPeter StahlDetlef von Hoerschelmann
    • Albert RoderHans MolleringWolfgang GruberKlaus BeaucampHans SeidelPeter StahlDetlef von Hoerschelmann
    • C12N9/08C12Q1/28G01N33/50C07G7/02C12D13/10G01N31/14
    • C12N9/0065C12Q1/28
    • 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.Mto hydrogen peroxide of 2.8.times.10.sup.-5 M andto 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.
    • 提供了一种用于还原型烟酰胺 - 腺嘌呤二核苷酸的新型过氧化物酶,其过氧化物酶用过氧化氢氧化还原型烟酰胺 - 腺嘌呤二核苷酸,得到烟酰胺 - 腺嘌呤二核苷酸和水,其特征在于具有2.8x10-5M的过氧化氢的米氏常数KM, 降低了1.7×10-5M的烟酰胺 - 腺嘌呤二核苷酸,在25℃下,在含有0.1M醋酸钾的0.2M Tris缓冲液pH6.0中测定。 过氧化物酶可以通过消化或通过用表面活性剂处理从粪链球菌ATCC 8043中释放酶制备,并从获得的酶溶液中分离。 过氧化氢在简单的反应或与特定氧化酶的偶联反应中通过使所述过氧化物酶与产生H 2 O 2的反应混合物在6.0至9.0的pH下接触并测定作为H 2 O 2和 最初存在于所述反应混合物中的烟酰胺 - 腺嘌呤二核苷酸的浓度。