Title: Spectrometric assay for horseradish peroxidase activity based on the linkage of conjugated system formed by oxidative decarboxylation
Abstract: Horseradish peroxidase (HRP)-catalyzed oxidation of 2,2-bis[3-acethylfilicinic acid-5-yl]acetic acid (BAFA, 4) produces Dehydro-3,3’-diacetyl-5,5’-methylenedifilicinic acid (DDMF, 3). A new photometric hydrogen donor (4) for peroxidase (POD)-catalyzed oxidation was demonstrated to be potentially useful for spectrocolorimetric and spectrofluorometric determination of HRP. Our developed colorimetric (absorption at 483 nm) and fluorometric (emission at 529 nm) systems both gave a linear calibration curve for HRP (y = 0.0025 × + 0.0237; R2 = 0.9997, and y = 0.241 × + 3.194; R2 = 0.9914, respectively) in the same concentration range of 9.1 × 10− 8–1.1 × 10− 6 nmol/L. The calculated Km and Vmax values of 5.10 × 10− 4 M and 5.13 × 10− 6 M/min, respectively. The results indicate that the quantification of HRP using BAFA 4 as hydrogen donor is possible.
Publication Year: 2016
Publication Date: 2016-11-13
Language: en
Type: article
Indexed In: ['crossref', 'pubmed']
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Cited By Count: 2
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