在有氧条件下,利用DEAE Sepharose Fast Flow弱阴离子离子交换层析和Blue Sepharose CL-6B亲和层析,同时分离并提纯克雷伯杆菌胞内的1,3-丙二醇氧化还原酶和甘油脱氢酶.研究表明,1,3-丙二醇氧化还原酶的纯化倍数为35.86倍,回收率为5.17%,该酶最适表观反应温度为57℃,最适反应pH值为9.5.在30℃及pH=8.0~10.0时,该酶具有良好的稳定性.在45℃和pH=9.5条件下,该酶以1,3-丙二醇和NAD+为底物,其米氏常数Km分别为15.8,0.2 mmol.L-1.1,3-丙二醇氧化还原酶对生理反应底物3-羟基丙醛活性最大,对其他醇类也有氧化能力.Mn2+对酶有显著激活作用,巯基保护剂能明显提高酶的活力.
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