Castillo M P, Stenström J, Ander P
Swedish University of Agricultural Sciences, Department of Microbiology, Uppsala.
Anal Biochem. 1994 May 1;218(2):399-404. doi: 10.1006/abio.1994.1198.
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.
这种方法早些时候被提出用于在过氧化物酶 - 葡萄糖氧化酶系统中测量葡萄糖,但尚未用于测定锰过氧化物酶(MnP)的活性。该测定基于3 - 甲基 - 2 - 苯并噻唑啉酮腙(MBTH)和3 - (二甲基氨基)苯甲酸(DMAB)的氧化偶联。在H2O2、Mn2 +和MnP存在下,MBTH与DMAB的反应产生深紫蓝色,具有宽吸收带,峰值在590nm处。消光系数很高(53,000 M-1 cm-1),因此可以检测到低水平的MnP活性。通常存在于白腐真菌培养物中的木质素过氧化物酶和漆酶,在所测试的浓度下几乎没有干扰或没有干扰。然而,在极低的MnP活性下,非常高的LiP活性可能会产生轻微干扰。