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用阶段密度梯度超速离心法,从蟾蜍血清中分离出7个组份,somanase(A酯酶)的活性在第3组份(35 ml离心管),或分成10个组分,则活性在第5组份(10 ml离心管),提纯12倍,对重复加入的梭曼水解较快。用苏丹黑预染聚丙烯酰胺凝胶电泳法,油红“0”脂蛋白染色琼脂糖电泳法,均显示脂蛋白染色呈阳性反应的一条区带,并求出K_m,V_(max)和相关系数值。在含蔗糖—KBr的介质中稳定性好,60℃3d未失活。对脂质部分的定量分析、分子量测定、电镜检查、及飘浮系数值均符合HDL的特征,因此,血清中的梭曼水解酶是HDL的一种成分。
Seven components were isolated from toad serum by stage density gradient ultracentrifugation. The activity of somanase (A-esterase) was in fraction 3 (35 ml centrifuge tubes) or divided into 10 fractions. The activity of 5 fractions (10 ml centrifuge tube), purify 12 times and hydrolyze Soman with repeated addition. Sudan black pre-stained polyacrylamide gel electrophoresis, oil red “0” lipoprotein staining agarose electrophoresis, showed a positive lipoprotein staining zone, and find K_m, Vmax and related Coefficient value. The stability in sucrose-KBr medium was good, and it was not inactivated at 60 ℃ for 3d. Quantitative analysis of the lipid fraction, molecular weight measurement, electron microscopy, and floating coefficient values are in line with the characteristics of HDL, so the serum of soman hydrolase is a component of HDL.