Purification of plasmalogens using Rhizopus delemar lipase and Naja naja naja phospholipase A2.
Purification of plasmalogens using Rhizopus delemar lipase and Naja naja naja phospholipase A2.
复制标题
使用Rhizopus delemar 脂肪酶和Naja naja naja 磷脂酶A2 纯化缩醛磷脂。
作者：
Hirashima,Y;Farooqui,AA;Murphy,EJ;Horrocks,LA
牛心ChoGpl(胆碱甘油磷脂)和牛脑EtnGpl(乙醇胺甘油磷脂)含有二酰基、烯基和烷基类似物。用R.实现了血浆原的纯化。脱脂酶和n。眼镜蛇磷脂酶A2消化。在那里。脱脂酶可使1，2-二酰基甘油磷脂1-位上的酰基键发生水解性反应。然后。眼镜蛇磷脂酶A2对二酰基和烷基磷脂的活性大于对烯基酰基甘油磷脂的活性。这些酶主要用于脱除二酰基和烷基类似物。当双酰基类型通过与R.在脂肪酶中，血浆蛋白原含量分别为94.2%±0.21%(平均±0.21%，n=4)和94.9%±0.19%(平均±0.19%，n=3)。回收率分别为74%和88%。这些部分纯化的血浆蛋白原用N处理。眼镜蛇磷脂酶A2。PLsCHO和PlsEtn的血浆原纯度分别为97.7%±0.24%(Mean±S.E.M.，n=4)和98.8%±0.27%(Mean±S.E.M.，n=3)。血浆原回收率分别为7.7%±0.7%(Mean±S.E.M.，n=4)和10.2%±1.2%(Mean±S.E.M.，n=3)。
Bovine heart ChoGpl (choline glycerophospholipid) and bovine brain EtnGpl (ethanolamine glycerophospholipid) contain diacyl, alkenylacyl and alkylacyl analogs. Purification of plasmalogens was achieved usingR. delemarlipase andN. naja najaphospholipase A2digestion. TheR. delemarlipase hydrolyzes the acyl bond at the 1‐position of 1,2‐diacyl glycerophospholipids. TheN. naja najaphospholipase A2has greater activity with diacyl and alkylacyl than with alkenylacyl glycerophospholipids. These enzymes were mainly used to remove diacyl and alkylacyl analogs respectively. When the diacyl types were removed by double incubation withR. delemar lipase, the plasmalogen content was 94.2%±0.21% (mean±S.E.M., n=4) for PlsCho (plasmenylcholine) and 94.9%±0.19% (mean±S.E.M., n=3) for PlsEtn (plasmenylethanolamine). Recoveries were 74% and 88% respectively. These partially purified plasmalogens were treated withN. naja najaphospholipase A2. Finally, 97.7%±0.24% (mean±S.E.M., n=4) and 98.8%±0.27% (mean±S.E.M., n=3) pure plasmalogens were obtained for PlsCho and PlsEtn respectively. Plasmalogens were recovered in an overall yield of 7.7%±0.7% (mean±S.E.M., n=4) and 10.2%±1.2% (mean±S.E.M., n=3) for PlsCho and PlsEtn.