NADH Peroxidase for hydrogen peroxide determination

US4186052A · kind A · utility

Assignee

• Boehringer Mannheim GmbH · DE

Inventors

• Albert Roder · Seeshaupt, DE
• Hans Mollering · Tutzing, DE
• Wolfgang Gruber · Bischofswerda, DE
• Klaus Beaucamp · Tutzing, DE
• Hans Seidel · Tutzing, DE
• Peter Stahl · New York, US
• Detlef von Hoerschelmann · Wielenbach, DE

Key dates

• Filing date: Jun 29, 1977
• Grant date: Jan 29, 1980
• Expiry date: Jun 29, 1997

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/28
• WIPO field: Biotechnology
• WIPO sector: Chemistry

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 PA1 to hydrogen peroxide of 2.8.times.10.sup.-5 M and PA1 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.