磷脂酰-N-乙酰氨基葡萄糖苷的合成及脂质体制备

Synthesis and Liposome Preparation of Phosphatidyl-N-Acetylglucosamine

为了提升磷脂酰-N-乙酰氨基葡萄糖苷(PtdGlcNAc)的合成效率,选取不同的咪唑类离子液体,通过以磷脂酶D(phospholipase D,PLD)催化磷脂酰胆碱(phosphatidylcholine,PC)与N-乙酰-D-氨基葡萄糖(N-acetyl-beta-D-glucosamine,GlcNAc)的转酯反应,探究离子液体预处理对酶催化性能的影响。结果表明,经过1-丁基-3-甲基咪唑六氟磷酸盐([BMIm][PF_(6)])预处理后,PLD的活性明显高于其他离子液体组和空白组。因此,采用[BMIm][PF_(6)]作为PLD预处理的孵育溶液,通过优化实验,得到最佳反应条件为:1 U的加酶量,底物PC与GlcNAc的物质的量比为1:60,环戊基甲醚作为有机相,离子液体与水体积比为4:1,反应时间12 h,产率为79.7%,与未处理组的产率相比提升了56.9%。将合成的PtdGlcNAc制备成纳米脂质体,并对其结构进行了初步表征。该研究丰富了PLD反应体系的相关研究,为其他新型磷脂酰化合物的合成和应用提供了参考。

The effect of ionic liquid pretreatment on the enzyme catalytic activity was investigated to improve the synthesis efficiency of phosphatidyl-N-acetylglucosamine(PtdGlcNAc)and different imidazole ionic liquids were selected to study the transphosphatidylation of phosphatidylcholine(PC)and N-acetyl-beta-D-glucosamine(GlcNAc)catalyzed by phospholipase D(PLD).The results showed that the enzyme activity of PLD pretreated with 1-butyl-3-methylimidazole hexafluorophosphate([BMIm][PF_(6)])was significantly higher than that of other ionic liquids and untreated blank groups.Therefore,[BMIm][PF_(6)]was used as the incubation solution for PLD pretreatment.Through the optimization experiment,the optimum reaction conditions were as follows:1 U PLD,the amount ratio of substrate PC to GlcNAc of 1∶60,cyclopentyl methyl ether as the organic phase,and the volume ratio of ionic liquid phase to aqueous phase of 4∶1.After 12 h,the product yield was 79.7%,which was 56.9%higher than that of the untreated one.PtdGlcNAc were synthesized into nanoliposomes which structures were preliminary characterized.This study could enrich the related research of PLD and provide a reference for the synthesis and application of other novel phosphatidyl compounds.