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Journal of the Korean Chemical Society (JKCS)

ISSN 1017-2548(Print)
ISSN 2234-8530(Online)
Volume 37, Number 10
JKCSEZ 37(10)
August 20, 1993 

Analytical Studies of H2O2-Producing Oxidase Systems

H2O2-생성 산화효소계에 관한 분석 연구
Younghee Hahn, Hae-Lim Cho

한영희, 조혜림
산소를 수용체로 하여 H2O2를 생성하는 산화효소인 glucose oxidase (GO) 와 alcohol oxidase (AO)를 이용하여 glucose와 ethanol에 대한 분광학적 효소분석법과 전기화학적 효소분석법을 연구하였다. Glucose에 대한 정량분석의 경우 GO 반응에서 생성된 H2O2를 peroxidase를 사용하여 K4Fe(CN)6에 반응시키고 그 결과 생성된 K3Fe(CN)6의 흡광도를 418 nm에서 측정하거나 Ag/AgCl (포화 KCl) 기준전극에 대하여 -55 mV를 부하시킨 유리질 탄소전극에서 K3Fe(CN)6의 확산전류를 측정하였다. 전류법 측정이 분광광도법 측정보다 1/1000 정도 더 낮은 농도까지 검출할 수 있었으며 직선성을 갖는 농도 범위도 10배 더 연장되었다. Ethanol에 대한 정량분석의 경우 K3Fe(CN)6가 AO에 의하여 순간적으로 소거되었으므로 AO 반응만을 이용하여 H2O2의 생성속도를 +0.900 V에서 측정하거나 용존 O2의 감소속도를 -0.500 V에서 전류법으로 측정하였다.

Spectrophotometric enzymatic analysis and amperometric enzymatic analysis for the determinations of glucose and ethanol were studied utilizing glucose oxidase (GO) and alcohol oxidase (AO), respectively, which commonly consume O2 and produce H2O2. For the determination of glucose, H2O2 were coupled to K4Fe(CN)6 via peroxidase producing K3Fe(CN)6 whose absorbance was measured at 418 nm or whose diffusion current was measured on the glassy carbon electrode at an applied potential of -55 mV vs. Ag/AgCl (sat. KCl) reference electrode. Amperometric analysis was 1000 times more sensitive as well as 10 times better in the linear concentration range than spectrophotometric analysis. For the determination of ethanol, AO only was used for the enzymatic analysis, since K3Fe(CN)6 was completely disappeared as soon as AO was added. Either rate of H2O2 produced was amperometrically measured at +0.900 V or rate of O2 consumed was measured at -0.500 V vs. Ag/AgCl(sat. KCl) reference electrode.

874 - 880
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