Production of a Novel Protopanaxatriol-Type Ginsenoside by Yeast Cell Factories

Abstract

Ginsenosides, the main active compounds in Panax species, are glycosides of protopanaxadiol (PPD) or protopanaxatriol (PPT). PPT-type ginsenosides have unique pharmacological activities on the central nervous system and cardiovascular system. As an unnatural ginsenoside, 3,12-Di-O-β-D-glucopyranosyl-dammar-24-ene-3β,6α,12β,20S-tetraol (3β,12β-Di-O-Glc-PPT) can be synthesized through enzymatic reactions but is limited by the expensive substrates and low catalytic efficiency. In the present study, we successfully produced 3β,12β-Di-O-Glc-PPT in Saccharomyces cerevisiae with a titer of 7.0 mg/L by expressing protopanaxatriol synthase (PPTS) from Panax ginseng and UGT109A1 from Bacillus subtilis in PPD-producing yeast. Then, we modified this engineered strain by replacing UGT109A1 with its mutant UGT109A1-K73A, overexpressing the cytochrome P450 reductase ATR2 from Arabidopsis thaliana and the key enzymes of UDP-glucose biosynthesis to increase the production of 3β,12β-Di-O-Glc-PPT, although these strategies did not show any positive effect on the yield of 3β,12β-Di-O-Glc-PPT. However, the unnatural ginsenoside 3β,12β-Di-O-Glc-PPT was produced in this study by constructing its biosynthetic pathway in yeast. To the best of our knowledge, this is the first report of producing 3β,12β-Di-O-Glc-PPT through yeast cell factories. Our work provides a viable route for the production of 3β,12β-Di-O-Glc-PPT, which lays a foundation for drug research and development.

A systematic review of ginsenoside biosynthesis, spatiotemporal distribution, and response to biotic and abiotic factors in American ginseng

“…(2) Metabolic engineering of plant specialized metabolites in microorganisms or plant chassis could greatly improve the production of valuable ginsenosides. 170,171 Green bioproduction of few ginsenosides, such as Rh1, 172 Rh2, 173 CK, 174 and F1, 175 has been achieved based on the biosynthetic pathway from P. ginseng . However, the biosynthesis of many ginsenosides remains unclear, and metabolic engineering faces challenges, including novel gene element identification, chassis selection and modification, and fermentation engineering optimization, to improve the titers of the target ginsenosides.…”

Therapeutic Applications of Ginseng Natural Compounds for Health Management

“…Thus, the literature shows that PPD prevents and is a promising treatment for endometriosis [29]. On the other hand, PPT ginsenosides have pharmacological effects on the central nervous and cardiovascular systems [30], and PPT reportedly acts as a PPARγ antagonist [31]; thus, targeting PPARγ is considered a promising treatment option for obesity.…”