三七茎叶总皂苷与I型人参皂苷酶反应的动态变化
Dynamic changes of multi-notoginseng stem-leaf ginsenosides in reaction with ginsenosidase type-I.
作者信息
Xiao Yongkun, Liu Chunying, Im Wan-Teak, Chen Shuang, Zuo Kangze, Yu Hongshan, Song Jianguo, Xu Longquan, Yi Tea-Hoo, Jin Fengxie
机构信息
Department of Oriental Medicinal Biotechnology, College of Life Science, Kyung Hee University, Global Campus, Yongin, Republic of Korea.
College of Biotechnology, Dalian Polytechnic University, Dalian, China.
出版信息
J Ginseng Res. 2019 Apr;43(2):186-195. doi: 10.1016/j.jgr.2017.10.001. Epub 2017 Oct 21.
BACKGROUND
Notoginseng stem-leaf (NGL) ginsenosides have not been well used. To improve their utilization, the biotransformation of NGL ginsenosides was studied using ginsenosidase type-I from g.848.
METHODS
NGL ginsenosides were reacted with a crude enzyme in the RAT-5D bioreactor, and the dynamic changes of multi-ginsenosides of NGL were recognized by HPLC. The reaction products were separated using a silica gel column and identified by HPLC and NMR.
RESULTS
All the NGL ginsenosides are protopanaxadiol-type ginsenosides; the main ginsenoside contents are 27.1% Rb3, 15.7% C-Mx1, 13.8% Rc, 11.1% Fc, 7.10% Fa, 6.44% C-Mc, 5.08% Rb2, and 4.31% Rb1. In the reaction of NGL ginsenosides with crude enzyme, the main reaction of Rb3 and C-Mx1 occurred through Rb3→C-Mx1→C-Mx; when reacted for 1 h, Rb3 decreased from 27.1% to 9.82 %, C-Mx1 increased from 15.5% to 32.3%, C-Mx was produced to 6.46%, finally into C-Mx and a small amount of C-K. When reacted for 1.5 h, all the Rb1, Rd, and Gyp17 were completely reacted, and the reaction intermediate F2 was produced to 8.25%, finally into C-K. The main reaction of Rc (13.8%) occurred through Rc→C-Mc1→C-Mc→C-K. The enzyme barely hydrolyzed the terminal xyloside on 3-- or 20--sugar-moiety of the substrate; therefore, 9.43 g C-Mx, 6.85 g C-K, 4.50 g R7, and 4.71 g Fc (hardly separating from the substrate) were obtained from 50 g NGL ginsenosides by the crude enzyme reaction.
CONCLUSION
Four monomer ginsenosides were successfully produced and separated from NGL ginsenosides by the enzyme reaction.
背景
三七茎叶（NGL）皂苷尚未得到充分利用。为提高其利用率，利用来自 g.848 的 I 型人参皂苷酶研究了 NGL 皂苷的生物转化。
方法
NGL 皂苷在 RAT-5D 生物反应器中与粗酶反应，通过高效液相色谱（HPLC）识别 NGL 多种皂苷的动态变化。反应产物用硅胶柱分离，通过 HPLC 和核磁共振（NMR）鉴定。
结果
所有 NGL 皂苷均为原人参二醇型皂苷；主要皂苷含量分别为：Rb3 27.1%、C-Mx1 15.7%、Rc 13.8%、Fc 11.1%、Fa 7.10%、C-Mc 6.44%、Rb2 5.08%、Rb1 4.31%。在 NGL 皂苷与粗酶的反应中，Rb3 和 C-Mx1 的主要反应过程为 Rb3→C-Mx1→C-Mx；反应 1 小时后，Rb3 从 27.1%降至 9.82%，C-Mx1 从 15.5%增至 32.3%，生成 C-Mx 至 6.46%，最终转化为 C-Mx 和少量 C-K。反应 1.5 小时后，所有 Rb1、Rd 和 Gyp17 完全反应，生成反应中间体 F2 至 8.25%，最终转化为 C-K。Rc（13.8%）的主要反应过程为 Rc→C-Mc1→C-Mc→C-K。该酶几乎不水解底物 3-或 20-糖基上的末端木糖苷；因此，通过粗酶反应从 50 g NGL 皂苷中获得了 9.43 g C-Mx、6.85 g C-K、4.50 g R7 和 4.71 g Fc（几乎无法与底物分离）。
结论
通过酶反应成功从 NGL 皂苷中制备并分离出四种单体皂苷。
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