原人参二醇类皂苷(PPD)在酶反应中转化动态及其产物稀有皂苷的制备

Protopanaxadiol ginsenosides dynamics in enzymatic reaction and preparation of rare ginsenosides

目的为了生物转化低成本制备人参稀有皂苷,利用Aspergillus g.848菌的粗酶与市售的原人参二醇类皂苷(PPD)混合皂苷反应,制备人参稀有皂苷C-K、C-Mc、F_2单体和4种异构体的Rh2组皂苷。方法酶与PPD皂苷反应,生成稀有皂苷;用HPLC法测定PPD皂苷原料和产物皂苷的组成,用硅胶柱法分离产物中的单体皂苷,用NMR法确认产物单体皂苷结构;用UPLC-MS法确认产物Rh2组。结果原料PPD中含有人参皂苷Rb_1、Rd、Rb_2、Rc和4种异构体的人参皂苷Rg3组;酶反应时,若生产以F_2为主的皂苷时,最佳反应时间为1.5~2.0h;若生产以C-K为主的皂苷时,反应时间为24.0~30.0h;若生产以Rh2组为主的皂苷时,反应6.0~12.0h时,Rg_3组产量低而Rh_2组产量高。以C-K为主的皂苷生产中,从30 g的PPD皂苷酶反应得到20 g产物,经硅胶柱分离,得到8.16 g的C-K、1.01 g的C-Mc、0.45 g的F_2单体和0.19 g的Rh_2组皂苷,并以NMR和UPLC-MS法核对了其结构。结论 PPD皂苷经酶转化成功地制备了高活性C-K、C-Mc、F_2和Rh_2组皂苷。

Objective To cheaply prepare the rare ginsenosides by biotransformation, ginsenosides C-K, C-Mc, F2, and Rh2 from commercially available Protopanaxadiol （PPD） ginsenoside mixture were prepared using a crude enzyme of Aspergillus g.848 strain. Methods The rare ginsenosides were obtained from PPD ginsenosides by enzyme reaction; The composition of PPD raw materials and ginsenoside products was measured by HPLC. The monomer ginsenosides and Rh2 group of enzyme reaction product were separated by a silica gel column; The produced monomer ginsenosides were identified by NMR; Rh2 group was identified by UPLC-MS. Results The raw material ofPPD ginsenosides was consisted ofginsenoside Rbb Rd, Rb2, Rc, and Rg3 groups with four kinds of isomers. During the reaction of enzyme, the best reaction time for the ginsenoside F2 production was 1.5 h to 2 h; The best reaction time for the ginsenoside C-K production was 24 h to 30 h; If producing the Rh2 group, when reacted to 6 h to 12 h, the content of Rg3 group was low, and Rh2 group was high, In the production of C-K, 20 g of crude products were obtained from 30 g of PPD ginsenosides by enzyme reaction, and 8.16 g of C-K, 1.01 g of C-MC, 0.45 g ofF2, and 0.19 g of Rh2 group were separated using silica gel column. The rare ginsenosides were identified by NMR, and the RhE group was identified using UPLC-MS method. Conclusion The high activity of monomer ginsenoside C-K, C-Mc, F2, and Rh2 group are successfully prepared from the PPD ginsenosides by enzymatic conversion.