2-花生四烯酸单甘酯(2-AG)是一种内源性大麻素,在神经、心血管和免疫等系统中具有一系列的生理活性.为实现2-AG的绿色高效制备,探究了脂肪酶催化醇解富含花生四烯酸的微生物油制备2-AG的方法.以2-单甘酯(2-MAG)含量为指标,通过单因素实验对酶法催化醇解反应条件进行了优化,并采用溶剂萃取法对产物进行纯化.结果表明:最佳反应条件为以Lipozyme 435脂肪酶为醇解脂肪酶、酶添加量4%(以油质量计)、油与无水乙醇物质的量比1∶40、叔丁醇为溶剂、油溶比2∶3、反应温度35℃、反应时间8 h,在最佳条件下粗产物中2-MAG含量为33.63%;经溶剂萃取纯化后2-MAG的纯度达到了94.79%,其中2-AG含量为40.70%.综上,酶法醇解富含花生四烯酸的微生物油可以获得高2-AG含量的2-MAG.
2-Arachidonoylglycerol(2-AG)is an endogenous cannabinoid and exhibits a variety of biological activities in the central nervous,cardiovascular,and immune systems.To prepare 2-AG in a greener and efficient way,a lipase-catalyzed alcoholysis of microbial oil rich in arachidonic acid was used to synthesize 2-AG.With the 2-monoacylglycerol(2-MAG)content as an index,the reaction conditions of lipase-catalyzed alcoholysis were optimized by single factor experiment,and a solvent extraction method was employed to purify the product.The results showed that the optimal reaction conditions of lipase-catalyzed alcoholysis were as follows:Lipozyme 435 as alcoholysis enzyme,lipase dosage 4%(relative to oil mass),molar ratio of oil to anhydrous ethanol 1∶40,tert-butanol as solvent,ratio of oil to solvent 2∶3,reaction temperature 35 ℃ and reaction time 8 h.Under the optimal conditions,2-MAG content in the crude product was up to 33.63%.The purity of 2-MAG after purification was up to 94.79%,and the content of 2-AG was 40.70%.In conclusion,enzymatic alcoholysis of microbial oil rich in arachidonic acid can obtain 2-MAG with high 2-AG content.