一种利用蜗牛酶转化重楼皂苷Ⅵ制备偏诺皂苷元的方法  

A Method for Preparation of Pennogenin by Converting Polyphyllin VI with Snailase

作　　者：王亢宗 李敏 田济铭 张嘉颀 薛哲勇 宋伟 华欣 Wang Kangzong;Li Min;Tian Jiming;Zhang Jiaqi;Xue Zheyong;Song Wei;Hua Xin(School of Life Sciences,Northeast Forestry University,Harbin 150006,China)

机构地区：[1]东北林业大学生命科学学院,哈尔滨150006

出　　处：《世界科学技术-中医药现代化》2022年第5期1760-1771,共12页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：中国中医科学院中央本级重大增减支项目“名贵中药资源可持续利用能力建设项目”(2060302-2101-17):重楼皂苷代谢途径解析,负责人:薛哲勇。

摘　　要：目的 建立并优化一种利用蜗牛酶水解重楼皂苷Ⅵ制备偏诺皂苷元的转化方法。方法 利用体外酶促体系,使用蜗牛酶水解重楼皂苷Ⅵ制备偏诺皂苷元,通过高效液相色谱和核磁检测分离并鉴定产物结构。设计反应时间、酶用量、反应温度和pH值四个单因素,以转化率为考察指标,通过多项式回归及响应面法确定最优的反应条件。结果 通过单因素与正交实验,确定了实验室制备偏诺皂苷元的最优反应条件:反应温度为37°C、酶用量为1.5 mg(每100μg底物),反应时间36 h,pH为5.35,模型预测产率为53.5%,验证产率为51.5%,与预期基本相符。结论 开发并优化了一种用蜗牛酶转化重楼皂苷Ⅵ制备偏诺皂苷元的方法,该方法可行性高,反应得率高,耗时短,成本低,可能对开展中试研究具有潜在的指导作用。Objective To establish a conversion system of snailase hydrolyzing polyphyllin VI to prepare pennogenin.Methods Polyphyllin VI was hydrolyzed to pennogenin with snailase in vitro. The structure of the product was separated and identified by high performance liquid chromatography and nuclear magnetic resonance. The effectiveness of reaction time, enzyme dosage, temperature, and pH value were investigated, with conversion rate used as the inspection index to determine the optimal reaction conditions using polynomial regression and the response surface method.Results The optimal reaction conditions for the preparation of pennogenin in laboratory were determined using single factor and orthogonal experiments: reaction temperature was 37°C, enzyme dosage was 1.5 mg(reaction system 300 μL), reaction time was 36 hours, pH was 5.35. The predicted yield of the model was 53.5%, and the verified yield was 51.5%, both of which were largely expected. Conclusion It is a feasible way for converting polyphyllin VI into pennogenin using snailase, with a high reaction yield, a short time-consuming process, and a low cost, which may be instructive for conducting pilot tests.

关 键 词：重楼皂苷Ⅵ 偏诺皂苷元 蜗牛酶 多项式回归 

分 类 号：R283[医药卫生—中药学]