Studies on the enzymatic synthesis of a new PPT-type ginsenoside via UDP-glycosyltransferase PgUGT74AE2 from Panax ginseng
10.16438/j.0513-4870.2018-0300
- VernacularTitle:人参糖基转移酶PgUGT74AE2催化生成新型人参三醇皂苷研究
- Author:
Ting-ting ZHANG
1
;
Hui-chao LIANG
1
;
Ting GONG
1
;
Zong-feng HU
1
;
An-di GU
1
;
Jin-ling YANG
1
;
Ping ZHU
1
Author Information
1. State Key Laboratory of Bioactive Substance and Function of Natural Medicines and Key Laboratory of Biosynthesis of Natural Products of National Health Commission of PRC, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
- Publication Type:ORIGINAL ARTICLES
- Keywords:
Panax ginseng;
glycosyltransferase;
glycosylation;
protopanaxatriol;
protopanaxatriol-type ginsenoside
- From:
Acta Pharmaceutica Sinica
2018;53(9):1565-1570
- CountryChina
- Language:Chinese
-
Abstract:
UDP-glycosyltransferase PgUGT74AE2 from Panax ginseng can transfer a glucose moiety to the free C-3 hydroxyl of protopanaxadiol (PPD) to produce ginsenoside Rh2. However, no report demonstrates that PgUGT74AE2 can transfer a glucose moiety to the free C-3 hydroxyl of protopanaxatriol (PPT) to produce a PPT-type ginsenoside. In this study, the expression plasmid pET-32a-PgUGT74AE2 was constructed for expression of the recombinant protein and transferred into Escherichia coli Transetta (DE3) to generate the recombinant strain Transetta-PgUGT74AE2. The recombinant enzyme PgUGT74AE2 was expressed by induction of isopropyl-β-D-thiogalactoside (IPTG). An in vitro enzymatic reaction system was established with the recombinant enzyme PgUGT74AE2 and the substrate PPT. PgUGT74AE2 catalyzed the glycosylation of the free C-3 hydroxyl of PPT to produce 3-O-β-D-glucopyranosyl-dammar-24-ene-3β,6α,12β,20S-tetraol, a new PPT-type ginsenoside. This study provides an efficient approach for the biosynthesis of a new PPT-type ginsenoside through in vitro enzymatic reaction, which may pave a way to produce promising lead in drug discovery.
