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以中华猕猴桃鲜果为材料,经调节pH,硫酸铵盐析,除果胶以及DEAE—纤维素(E22)柱层折纯化,获得电泳单一区带酶制剂。以牛碳氧血红蛋白为底物,酶的最适温度为45—50℃,最适pH为3.7—4.0,K_m值为2.78×10~(-5)M,K_(cat)为2.01×10~2/min,pH影响V_(max)而不影响K_m值,表现为非竞争性抑制作用。该酶的活活化能为6.84kcal/mol,ΔH为6.24—6.20kcal/mol(30—50℃),同时还进行了酶的热稳定性研究。
Using Chinese kiwifruit fresh fruit as material, the electrophoresis single zone enzyme preparation was obtained by adjusting pH, ammonium sulfate salting out, removing pectin and DEAE-cellulose (E22) column chromatography. With bovine carboxyhemoglobin as the substrate, the optimum temperature of the enzyme was 45-50 ℃, the optimum pH was 3.7-4.0, K_m was 2.78 × 10 -5 M, K-cat was 2.01 × 10 ~ 2 / min, pH affected V max but did not affect K m, showing noncompetitive inhibition. The activation energy of the enzyme was 6.84 kcal / mol, the ΔH was 6.24-6.20 kcal / mol (30-50 ℃), and the thermal stability of the enzyme was also studied.