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. 2016 Oct;40(4):366-374.
doi: 10.1016/j.jgr.2015.11.004. Epub 2015 Dec 17.

Microbial Conversion of Major Ginsenosides in Ginseng Total Saponins by Platycodon grandiflorum Endophytes

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Free PMC article

Microbial Conversion of Major Ginsenosides in Ginseng Total Saponins by Platycodon grandiflorum Endophytes

Lei Cui et al. J Ginseng Res. .
Free PMC article

Abstract

Background: In this study, we screened and identified an endophyte JG09 having strong biocatalytic activity for ginsenosides from Platycodon grandiflorum, converted ginseng total saponins and ginsenoside monomers, determined the source of minor ginsenosides and the transformation pathways, and calculated the maximum production of minor ginsenosides for the conversion of ginsenoside Rb1 to assess the transformation activity of endophyte JG09.

Methods: The transformation of ginseng total saponins and ginsenoside monomers Rb1, Rb2, Rc, Rd, Rg1 into minor ginsenosides F2, C-K and Rh1 using endophyte JG09 isolated by an organizational separation method and Esculin-R2A agar assay, as well as the identification of transformed products via TLC and HPLC, were evaluated. Endophyte JG09 was identified through DNA sequencing and phylogenetic analysis.

Results: A total of 32 β-glucosidase-producing endophytes were screened out among the isolated 69 endophytes from P. grandiflorum. An endophyte bacteria JG09 identified as Luteibacter sp. effectively converted protopanaxadiol-type ginsenosides Rb1, Rb2, Rc, Rd into minor ginsenosides F2 and C-K, and converted protopanaxatriol-type ginsenoside Rg1 into minor ginsenoside Rh1. The transformation pathways of major ginsenosides by endophyte JG09 were as follows: Rb1→Rd→F2→C-K; Rb2→C-O→C-Y→C-K; Rc→C-Mc1→C-Mc→C-K; Rg1→Rh1. The maximum production rate of ginsenosides F2 and C-K reached 94.53% and 66.34%, respectively.

Conclusion: This is the first report about conversion of major ginsenosides into minor ginsenosides by fermentation with P. grandiflorum endophytes. The results of the study indicate endophyte JG09 would be a potential microbial source for obtaining minor ginsenosides.

Keywords: Platycodon grandiflorum; conversion; endophytes; minor ginsenosides.

Figures

Fig. 1
Fig. 1
Gel electrophoresis of recombinant plasmid DNA of endophyte JG09.
Fig. 2
Fig. 2
A 16S rDNA sequenced-based phylogenetic tree constructed by the Neighbor joining method. The numbers at the nodes are bootstrap values based on 1,000 replications.
Fig. 3
Fig. 3
TLC analysis of metabolites of ginseng total saponins by endophyte JG09 cultured in different media: H2O (1), R2A (2), PDB (3), LB (4), NB (5), and LL (6). LB, Luria-Bertani; LL, Lysogeny Liquid; NB, Nutrient Broth; PDB, Potato Dextrose Broth; R2A, Agar medium S; S1 and S2, saponin standards.
Fig. 4
Fig. 4
TLC analysis of metabolites of ginseng total saponins by endophyte JG09 cultured at different pH's of 3.0 (1), 4.0 (2), 5.0 (3), 6.0 (4), 7.0 (5), 8.0 (6), 9.0 (7), and 10.0 (8). S1 and S2, saponin standards.
Fig. 5
Fig. 5
HPLC analysis of (A) ginseng total saponins and (B) fermented ginseng total saponins. The peaks correspond to ginsenoside Rg1, Re, Rh1, Rb1, Rc, Rb2, Rd, F2, and C-K, respectively.
Fig. 6
Fig. 6
Time-course TLC analyses of ginsenoside (A) Rb1, (B) Rb2, (C) Rc, (D) Rd, and (E) Rg1 by endophyte JG09. C, blank sample; S1–S5, standard.
Fig. 7
Fig. 7
(A) HPLC profile of the metabolites of ginsenoside Rb1 converted by endophyte JG09. (B) The content change of ginsenoside Rb1, Rd, F2, and C-K in the fermentation broth.
Fig. 8
Fig. 8
Biotransformation pathway of ginsenosides Rb1, Rb2, Rc, and Rd by endophyte JG09.
Fig. 9
Fig. 9
Biotransformation pathway of ginsenosides Rg1 by endophyte JG09.
Fig. 10
Fig. 10
Standard curve of ginsenoside F2 and C-K.

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