Resazurin as an Electron Acceptor in Glucose Oxidase-Catalyzed Oxidation of Glucose

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The behavior of resazurin(1)as an electron acceptor in glucose oxidase(GOD)-catalyzed oxidation of glucose under anaerobic conditions is described. When a mixture of 1, glucose, and GOD in phosphate buffer(pH 7.4, 0.1M)was incubated at 25℃, the resulting solution turned purple to fluorescent pink due to the deoxygenated product, resorufin(2). On incubation of 1 with GOD alone or with H_2O_2 under essentially the same conditions, no color change was seen, indicating that generation of 2 in the enzymatic reaction is brought about through reduction of 1 by the reduced form(GOD_<red>)of GOD, which was also supported by the voltammetric behavior of 1. However, it was found that the enzymatic transformation of 1 to 2 is of no practical use as an indicator reaction for glucose determination using only GOD due to a slow reaction of 1 with GOD_<red>. Based on a pingpong type mechanism with a steady-state approximation, K_M and k_<cat> for 1 as an electron acceptor from GOD_<red> were estimated to be 15±1.3μM and (5.0±0.5)×10^<-2>s^<-1> respectively.
公益社団法人日本薬学会の論文
2001-05-01