胆碱脱氢酶与脂质体的重组

Reconstitution of Choline Dehydrogenase and Liposome

用预先制备的脂质体与含Triton X-100的胆碱脱氢酶保温随后用Bio-beadsSM-2去除去垢剂而使酶重组到脂质体,Triton X-100的浓度,磷脂-蛋白比,Mg^(2+),pH等均影响重组,脂质体的大小在重组中不是关键因素,重组酶的活力比纯化酶增加70％,重组酶对PMS的Km,相转变温度及活化能均与线粒体内膜接近而不同于纯化酶。

Choline dehydrogenase has been incorporated into liposome through a method that Triton X-100 was directly removed from liposome-detergent-enzyme solutions by Bic-beads SM-2. Concentration of Triton X-100, lipid/protein ratio, Mg2+, pH etc had influence on the reconstitution. Size of liposome wasn't a key factor in reconstitution procedure. Activity of reconstituted enzyme was increased 70% of that of purifed enzyme. Km for PMS, transition temperature and activation energy in reconstituted enzymes were similar to that in mitochondrial inner membrane,but were different from that in purifed enzyme.