维生素A-DHA酯的酶法合成及纯化制备  

Enzymatic synthesis and purification of Vitamin A docosahexaenoate

作　　者：高坤鹏 孙建安[1] 毛相朝[1,2] GAO Kunpeng;SUN Jian'an;MAO Xiangzhao(College of Food Science and Engineering,Ocean University of China,Qingdao 266003,China;Laboratory for Marine Drugs and Bioproducts of Pilot National Laboratory for Marine Science and Technology(Qingdao),Qingdao 266237,China)

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003 [2]青岛海洋科学与技术试点国家实验室海洋药物与生物制品功能实验室,山东青岛266237

出　　处：《南方水产科学》2022年第2期23-30,共8页South China Fisheries Science

基　　金：国家重点研发计划项目(2019YFD0901902)。

摘　　要：维生素A具有维持正常视力、改善皮肤、抗衰老等多种生理活性,但极不稳定,很容易在光、热、氧的作用下发生分解。选择二十二碳六烯酸乙酯(Ethyl docosahexaenoate,EDHE)与维生素A醋酸酯(Vitamin A acetate,VAAE)作为底物,在脂肪酶Novozyme435的催化下发生转酯反应,从而合成一种新型的维生素A-DHA酯(Vitamin A docosahexaenoate,VADHE)。利用液相及质谱对目的产物进行鉴定,在此基础上,利用硅胶柱层析(C18填料)对反应体系中的VADHE进行纯化。经液相色谱检测,VADHE的纯度达到90%以上。为了进一步提高目的化合物得率,分别在有机溶剂体系和无溶剂体系中对转酯反应进行了优化。在有机溶剂体系中,反应6 h转化率可达40.61%。在无溶剂体系中,用100 mg脂肪酶催化反应4 h,转化率可达56.39%。该研究首次设计并合成了新型脂质VADHE,实现了该物质的纯化制备,为后期的活性检测实验奠定了基础。Vitamin A shows a variety of physiological activities,such as maintaining normal vision,improving skin and antiaging activity.However,it is extremely unstable and easily decomposed under the action of light,heat and oxygen.In this study,Novozyme435 was used to catalyze the transesterification reaction of Vitamin A acetate(VAAE)and ethyl docosahexaenoate(EDHE).The target product(Vitamin A docosahexaenoate,VADHE)was identified by high performance liquid chromatography(HPLC)and mass spectrometry(MS).On this basis,VADHE in the reaction system was purified by silica gel column chromatography(C18 filler).The result of HPLC shows that the purity of VADHE reached 90%.In order to increase the yield of VADHE,the transesterification reaction was optimized in organic solvent system and solvent-free system,respectively.In the organic solvent system,the conversion rate reached 40.61%after 6 h.In the solvent-free system,when 100 mg Novozyme was used to catalyze the reaction for 4 h,the conversion rate reached 56.39%.VADHE was firstly designed and synthesized in this study,and the purified VADHE can be further used in the physiological activity experiment.

关 键 词：维生素A-DHA酯 脂肪酶 转酯反应 酶法合成 

分 类 号：Q814.9[生物学—生物工程]