발아중인 대두의 배에서 두 종류의 glyoxylate reductase(glycoIIate-NAD oxidoreductase, EC. 1. 1. 1. 26)를 추출하고 ammonium sulfate에 의한 침전, Sephadex G-200 gel filtration, DEAE-cellulose 및 Sephadex G-100 column chromatography에 의하여 정제하였다. 두 종류의 효소는 각각 NADH 및 NADPH를 cofactor로 요구하는 바 각각 745배 및 789배 정제하였다. 최종적으로 분리 정제된 상기 효소는 sucrose density gradient centrifugation, starch 및 acrylamide gel electrophoresis를 행하여 단일 효소 단백임을 확인하였다. NADH-linked glyoxylate reductase의 Michaelis constant, Km은 $9.0{ imes}10^{-3}M$ 이다. 또한 NADH-Iinked enzyme 및 NADPH-linked enzyme의 분자량은 Sephadex G-100 gel filtration으로 측정하여 각각 142,000 및 192,000 이었다. 이 효소의 최적 pH, 최적온도, thiol group 저해제 및 몇 가지의 금속 ion이 효소활성에 미치는 영향을 검토하였다.
Two protein fractions which catalyse the reduction of glyoxylate to glycollate were isolated from germinating soybean seed embryos by ammonium sulfate precipitation, Sephadex G-200 gel filtration. DEAE-cellulose column chromatography and rechromatography on Sephadex G-100. One protein fraction required NADH and another protein fraction required NADPH as cofactor. These two enzymes were purified 745-fold and 789-fold, respectively. The final preparations were homogeneous by sucrose density gradient centrifugation, starch and acrylamide gel electrophoresis. The apparent Michaelis constant. Km, of NADH-linked glyoxylate reductase was estimated as approximately $9.0{ imes}10^{-3}M$. Molecular weights of NADH-Iinked and NADPH-linked enzymes were estimated to be about 142,000 and 192,000, respectively, by Sephadex G-100 gel filtration. Effects of pH, temperature, thiol group inhibitors and various metal ions on the enzyme were also described.
