KSBB Journal

  • 제14권5호
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  • Pages.528-533
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  • 1999
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  • 1225-7117(pISSN)
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  • 2288-8268(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Acetobacter sp. CS5 Alcohol Dehydrogenase의 분리 및 특성

Purification and Characterization of Alcohol Dehydrogenase from Acetobacter sp. CS5

  • 발행 : 1999.10.01
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초록

초산생성 미생물인 분리균주 Acetobacter sp. CS5로부터 alcohol dehydrogenase(ADH)를 순수분리하였다. 이 효소는 Triton-X로 solubilization시킨 후 DEAE-Sephacel chromatography와 Sephacryl S-200 chromatography에 의해서 순수 분리되었다. 그 결과 15%의 수율과 14배의 정제된 효소를 얻었다. 정제된 효소의 분자량은 322 KDa으로 측정되었다. 또한 SDS-PAGE상에서는 분자량이 79 KDa, 49 KDa, 46KDa인 3개의 band를 확인하였고, 3분자의 79 KDa 소단위체와 각각 1분자인 49 KDa과 46 KDa인 소단위체로 이루어졌음을 확인하였다. 에탄올에 대한 Km값은 0.77 mM이었으며 최적 pH와 온도는 각각 4.0~5.0과 35$^{\circ}C$이였다.

Membrane-bound alcohol dehydrogenase(ADH) was purified to homogeneity from the acetic acid producing bacteria, Acetobacter sp. CS5. The enzyme was solubilized and extracted with Trition-X and purified using the DEAE-Sephacel chromatography and Sephacryl S-200 chromatography. The enzyme was purified to 14-fold with a yield of 15%. The molecular weight of the purified enzyme was to be 332 KDa. SDS-PAGE of the enzyme showed three subunits with molecular weights of 79 KDa, 49KDa and 46K Da. It indicated that enzyme consisted of three subunits of the 79 KDa, two subunits of the 49 KDa and. 46 KDa, respectively. The apparent Km value for ethanol was 0.77 mM and the optimum pH and temperature was 4.0-5.0 and 35$^{\circ}C$, respectively.

키워드

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