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Evaluation of an electrochemical method for the analysis of enzymatic inhibition reactions
1Faculty of Biotechnology, Fukui Prefectural University, Eiheiji, Fukui, Japan. kuematsu@fpu.ac.jp
Abstract:
An approximate equation for bioelectrocatalitic current was applied to an inhibition reaction analysis of bilirubin oxidase by anion (Cl(-), SCN(-), and F(-)) in order to assess the possibility of the electrochemical method for the analysis of enzymatic inhibition reactions. The approximate equation can be transformed into the Michaelis-Menten form, so that the bioelectrocatalitic current can be analyzed by the usual graphical manner, that is, Lineweaver-Burk, Hanes-Woolf, Dixon and Cornish-Bowden plots, if the rate of inhibition reaction is described as a simple Michaelis-Menten form. From the electrochemical assay, it was found that the inhibitor of Cl(-) and SCN(-) anions exhibit non-competitive inhibition while that of F(-) exhibits competitive inhibition, and their inhibition constants were 220, 45, and 22 mM, respectively. The results were essentially similar to those obtained from the conventional spectrophotometric assay.
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