Determination of manganese peroxidase activity with 3-methyl-2-benzothiazolinone hydrazone and 3-(dimethylamino)benzoic acid.
This method was proposed earlier for measuring glucose in a peroxidase-glucose oxidase system but has not been studied for determination of manganese peroxidase (MnP) activity. The assay is based on the oxidative coupling of 3-methyl-2-benzothiazolinone hydrazone (MBTH) and 3-(dimethylamino)benzoic acid (DMAB). The reaction of MBTH and DMAB in the presence of H2O2, Mn2+, and MnP gives a deep purple-blue color with a broad absorption band with a peak at 590 nm. The extinction coefficient is high (53,000 M-1 cm-1), so low MnP activities can be detected. Lignin peroxidase and laccase, usually present in cultures of white rot fungi, gave little or no interference at the concentrations tested. However, slight interference from very high LiP activity may occur at very low MnP activity.
Castillo
M P
MP
Swedish University of Agricultural Sciences, Department of Microbiology, Uppsala.
Stenström
J
J
Ander
P
P
D062365:Aminobenzoates / Q000378:metabolism*; D052160:Benzothiazoles; D002138:Calibration; D006835:Hydrazones; D006861:Hydrogen Peroxide; D007700:Kinetics; D008345:Manganese; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000378:metabolism*; D013053:Spectrophotometry; D013379:Substrate Specificity; D013844:Thiazoles / Q000378:metabolism*; D062368:meta-Aminobenzoates
D062365:Aminobenzoates; D052160:Benzothiazoles; D006835:Hydrazones; D013844:Thiazoles; D062368:meta-Aminobenzoates; D008345:Manganese; C028079:3-(dimethylamino)benzoic acid; D006861:Hydrogen Peroxide; D010544:Peroxidases; C042858:lignin peroxidase; C051129:manganese peroxidase; C002536:3-methyl-2-benzothiazolone hydrazone