嗜热糖苷酶Fpendo5A转化绞股蓝皂苷XLIX的产物鉴定及分离纯化  

Identification,isolation and purification of transformed product of gypenoside XLIX by thermophilic glycosidase Fpendo5A

作　　者：于渤浩 李男[1] 赵幻希 李晶[1] 郑飞[1] 戴雨霖 李卓[2] 刘淑莹[1,3] 于珊珊[1,4] YU Bo-hao LI Nan ZHAO Huan-xi LI Jing ZHENG Fei DAI Yu-lin LI Zhuo LIU Shu-ying YU Shan-shan(Changchun University of Chinese Medicine, Changchun 130000, Jilin Province, China)

机构地区：[1]长春中医药大学吉林省人参科学研究院,吉林长春130000 [2]长春中医药大学附属医院,吉林长春130021 [3]中国科学院长春应用化学研究所长春质谱中心,吉林长春130022 [4]上海交通大学生命科学技术学院,上海200240

出　　处：《中国生物制品学杂志》2017年第3期307-311,共5页Chinese Journal of Biologicals

基　　金：国家自然科学基金青年科学基金项目(31400682)

摘　　要：目的对嗜热糖苷酶Fpendo5A转化绞股蓝皂苷XLIX的产物进行鉴定及分离纯化。方法应用Fpendo5A酶转化绞股蓝皂苷XLIX,通过高效液相色谱法(HPLC)和快速分离液相色谱-四极杆-飞行时间质谱法(rapid resolution liquid chromatography-quadrupole-time of flight-mass spectrometry/mass spectrometry,RRLC-Q-TOF MS/MS)对转化产物进行鉴定,全制备型高效液相色谱对转化产物进行分离纯化。结果在65℃和p H 6.0条件下,利用Fpendo5A酶转化绞股蓝皂苷XLIX 48 h后,获得单一转化产物,该酶能够水解绞股蓝皂苷XLIX的C20位葡萄糖苷键,生成长梗绞股蓝皂苷Ⅰ。纯化后的转化产物纯度达95.42%,产率为21.6%。结论利用Fpendo5A酶能够成功转化绞股蓝皂苷XLIX,纯化后的绞股蓝皂苷转化产物纯度好,产率高,为今后的药理活性及相关基础研究奠定了基础。Objective To identify, isolate and purify the transformed product of gypenoside XLIX by thermophilic glycosidase FpendoSA. Methods Gypenoside XLIX was transformed with thermophilic glycosidase FpendoSA, and the transformed product was identified by HPLC, rapid resolution liquid chromatography-quadrupole-time of flight-mass spectrometry/mass spectrometry （RRLC-Q-TOF MS/MS）, then isolated and purified by fully preparative high perfor- mance liquid chromatography （HPLC）. Results Single product was obtained 48 h after transformation of gypenoside XLIX by thermophilic glycosidase FpendoSA at 65℃ and pH 6, which was gylongiposide I generated by hydrolysis of glycosidic bond at site C20 of gypenoside XLIX with FpendoSA. The purity and yield of the transformed product were 95. 42% and 21. 6% respectively after purification. Conclusion Gypenoside XLIX was successfully transformed by thermophilic glycosidase Fpendo5A. The purified transformed product reached a high purity and a high yield, which laid a foundation of further study on pharmacological activity as well as the relevant basic research.

关 键 词：绞股蓝皂苷 嗜热糖苷酶Fpendo5A 转化 高效液相色谱法 快速分离液相色谱-四极杆-飞行时间质谱法 全制备型高效液相色谱法 

分 类 号：O657.72[理学—分析化学]