| Title | Klebsiella pneumoniae 균주의 세포외막에서 분리한 2-furaldehyde dehydrogenase의 특성에 관한 연구 | ||
| Author | 이준우 · 강사욱 · 하영칠 · * 한홍의 | ||
| Address | 서울대학교 자연과학대학 미생물학과; * 인하대학교 이과대학 생물학과 | ||
| Bibliography | Korean Journal of Microbiology, 26(3),197-206, 1988 | ||
| DOI | |||
| Key Words | Klebsiella pneumoniae, 2-furaldehyde dehydrogenase, β-NAD^+, outer membrane | ||
| Abstract | An outer membrane-associated 2-furaldehyde dehydrogenase, catalyzing the oxidation of 2-furaldehyde to 2-furoic acid from Klebsiella pneumoniae was purified to homogeneity and characterized. The enzyme showed its highly specific dependency on β-NAD^+. Enzyme activity was monitored during purification by using substrate 2-furaldehyde and coenzyme β-NAD^+ by means of high performance liquid chromatography. The outer membrane was successfully collected by the methods of Percoll density gradient ultracentrifugation and ultracentrifugation after preferential solubilization of the membrane with Mg^2+ and Triton X-100. The enzyme was purified by the series of procedures including extraction of outer membrane protein with EDTA and lysozume, and fractionation by column chromatography on QAE-Sephades Q-50, and subsequently Sephadex G-100. The enzume showed its optimal activity at 85℃, pH 9.5, and in the presence of 1.5% (vol/vol) Triton X-100. The enzyme exhibited a native molecular size of 88,000 by nondenaturing polyacrylamide gel electrophoresis and had an apparent Km of 4.72mM for 2-furaldehyde. | ||
| Download PDF | Kor_260308_197-206p.pdf | ||