Title Klebsiella pneumoniae 균주의 세포외막으로부터 2-furaldehyde dehydrogenase의 부분정제에 관하여
Author 이준우 · 이병웅 · 강사욱 · 하영칠 · 유병설 * · 한홍의 **
Address 서울대학교 미생물학과; * 서울대학교 약학대학; ** 인하대학교 생물학과
Bibliography Korean Journal of Microbiology, 24(4),370-376, 1986
DOI
Key Words Klebsiella pneumoniae, 2-furaldehyde dehydrogenase, beta-nicotinamide adenine dinucleotide, high performance liquid chromatography
Abstract From the outer membrane portion of Gram-negative Klebsiella pneumoniae, the activity of 2-furaldehyde dehydrogenase depending upon beta-nicotinamide adenine dinucleotide was detected. Cytoplasmic membrane was preferentially extracted from crude membrane with Mg^2+ and Triton X-100, and then outer membrane was collected by ultracentrifugation. The crude enzyme was obtained by solubilization of outer membrane with lysozyme, ethylene diamine tetraacetate and Triton X-100. Thereafter 2-furaldehyde dehydrogenase was partially purified through column chromatography on QAE-Sephadex Q-50 and Sephadex G-150 and the enzyme activity was analyzed by means of high performance liquid chromatography. The optimal pH for the activity of the enzyme was about 9.5 and the optimal temperature was about 85℃. The partially purified enzyme retained tis activity at 85℃ for 5 hours. The optimal concentration of Triton X-100 for the activity of the enzyme was about 1.5% in the reaction mixture.
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