国产染料配基层析介质分离纯化甘油激酶α-磷酸甘油脱氢酶和心肌黄酶

PURIFICATION OF GLYCEROKINASE,α-GLYCEROPHOSPHATE DEHYDROGENASE AND DIAPHORASE BY DYE-LIGAND CHROMATOGRAPHY

以国产Sepharose 4 B和活性染料制备的YS一Ⅱ染料配基层析介质,分离提纯了甘油激酶、α-磷酸甘油脱氢酶和心肌黄酶,可使纯度分别提高6.6、2.9和3.3倍,收率为83.3、43.6和81.9%,所含的NADH 氧化酶和乳酸脱氢酶几乎全部可除去,各酶冻干后在10℃存放可稳定半年以上。所得的上述三个酶与自制的脂酶可组成血清甘油三酯酶联试剂盒。

YS-Ⅱ dye-ligand chromatographic media were prepared withdomestic materials and used satisfactorily to purify glycerokinase(1),a-glycero-phosphate dehydrogenase(2)and diaphorase(3).The purity of 1,2,and 3 increased6.6,2.9,and 3.3-fold,respectively.Their yields were 83.3,43.6,and 81.9%,respectively.Almost all of NADH oxidase(4)and lactic dehydrogenase(5)wereeliminated from the three enzymes.Lyophilized enzymes stored at 10℃ over 6months did not lose significant activity.The three enzymes combined with lipase(6)can be used in the diagnostic kit for testing the serum triglycerides level.