Purification and characterization of UDPG:ginsenoside Rd glucosyltransferase from suspended cells of Panax notoginseng

Yue, Cai-Jun; Zhong, Jian‐Jiang

doi:10.1016/j.procbio.2005.05.001

Cited by 52 publications

(26 citation statements)

References 32 publications

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“…Similarly, falcarinol, with concentrations ranging from 100 to 560 mg/ kg FW, was the major PA in all roots investigated (Table 1). This is also in accordance with previous investigations of American ginseng roots for ginsenosides and PAs [8,[34][35][36]43], although ginsenoside Rg 1 has been shown to be the major ginsenoside in some populations of American ginseng [11,33]. No significant differences in the relative concentrations of both ginsenosides and PAs were seen with increasing root weight (Fig.…”

Section: Resultssupporting

confidence: 92%

“…The glycosylation of triterpene aglycones proceeds via triterpene-glycosyltransferase enzymes, of which several candidates have been identified [3,42,43]. Based on the structures of the PPD-type ginsenosides, it appears that ginsenoside Rd is the precursor for ginsenoside Rb 1 , Rb 2 , Rb 3 , and Rc in ginseng roots, which is also in accordance with biosynthetic studies performed in cell cultures of Panax notoginseng [42,43] and in adventitious roots of transgenic P. ginseng [44]. Significant linear correlations between the concentrations of single ginsenosides of the PPD-type were found (Table 3), which confirms a close biosynthetic relationship among these ginsenosides.…”

Section: Correlation Between Ginsenosides and Pas In American Ginsengmentioning

confidence: 99%

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Biomass and content of ginsenosides and polyacetylenes in American ginseng roots can be increased without affecting the profile of bioactive compounds

Christensen

Jensen

2008

J Nat Med

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Fifty selected roots from a 7-year-old American ginseng (Panax quinquefolium L.) plant population grown in Denmark, with root weights varying from 191 to 490 g fresh weight (FW), were investigated for bioactive ginsenosides and polyacetylenes (PAs) in order to determine the correlation between the content of ginsenosides and PAs and root FW. PAs (falcarinol, panaxydol) and ginsenosides (Rb(1), Rb(2), Rb(3), Rc, Rd, Re, Rg(1)) were extracted from roots by sequential extraction with ethyl acetate and 80% methanol, respectively, and quantified in extracts by reverse-phase high-performance liquid chromatography (HPLC) using photodiode array detection. Total concentrations of PAs and ginsenosides varied between 150 and 780 mg/kg FW and 5,920 and 15,660 mg/kg FW, respectively. No correlation existed between the content of ginsenosides and PAs and root FW or between the total concentration of ginsenosides and PAs. Strong significant correlation was found between total content of ginsenosides and ginsenoside Rb(1) (r = 0.8190, P < 0.0001) and between total content of PAs and falcarinol (r = 0.9904, P < 0.0001). Based on the results of this study, it was concluded that it is possible to select large American ginseng roots for increased biomass production and concentration of bioactive ginsenosides and PAs without affecting the profile of bioactive compounds. Ginsenoside Rb(1) and falcarinol were found to be important selection parameters for identifying superior genotypes with the highest content of bioactive compounds.

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Section: Resultssupporting

confidence: 92%

Section: Correlation Between Ginsenosides and Pas In American Ginsengmentioning

confidence: 99%

Biomass and content of ginsenosides and polyacetylenes in American ginseng roots can be increased without affecting the profile of bioactive compounds

Christensen

Jensen

2008

J Nat Med

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“…On the other hand, we investigated whether the change in glucosyltransferase activity is regulated at the gene transcription level during elicitation with MJ. For glucosyltransferase activity assay in P. ginseng hairy root cultures, UGRdGT activity measurement was adopted as described by Yue and Zhong (2005). UGRdGT is an enzyme biotransfoming ginsenoside Rd to ginsenoside Rb1 in Panax notoginseng.…”

Section: Resultsmentioning

confidence: 99%

“…For the assay of UGRdGT activity, preparation of proteins was performed as described by Yue and Zhong (2005). Hairy roots (1 g) were ground in liquid nitrogen and resuspended in 2 ml of 50 mM Tris-HCl (pH 8.5), 2 mM EDTA, 10 mM 2-mercaptoethanol and 1 mM phenylmethylsulfonyl fluoride.…”

Section: Ugrdgt Activity Measurementmentioning

confidence: 99%

Upregulation of ginsenoside and gene expression related to triterpene biosynthesis in ginseng hairy root cultures elicited by methyl jasmonate

Kim

Bang

Kim

et al. 2009

Plant Cell Tiss Organ Cult

126

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In this study, methyl jasmonate (MJ)-induced changes of triterpene saponins in ginseng (Panax ginseng C.A. Meyer) hairy roots and expression profiling of relevant responsive genes were analyzed. The transcription of PgSS (squalene synthase), PgSE (squalene epoxidase), and PNA (dammarenediol synthase-II) genes in hairy root cultures elicited by MJ treatment increased as compared with the controls, whereas that of PNX (cycloartenol synthase) decreased slightly. In order to select candidate genes encoding for cytochrome P450-dependent hydroxylase or glucosyltransferase associated with triterpene biosynthesis, RT-PCR analysis was conducted following MJ elicitation. No differences were observed in any expression among the five genes associated with the cytochrome P450 family, when compared to that of control. For candidates of the glucosyltransferase gene,expression of EST IDs PG07020C06, PG07025D04, and PG07029G02 was upregulated. In an effort to assess the effects of MJ elicitation on the biosynthesis of triterpene saponin, protopanaxadiol saponin (Rb group) and protopanaxatriol saponin (Rg group) contents in hairy roots were evaluated by HPLC analysis. With 7 days of MJ elicitation, levels of all ginseonsides of the two-groups increased much higher than that observed in the control. In particular, protopanaxadiol-type saponin contents increased by 5.5-9.7 times that of the control, whereas protopanaxatriol-type saponin contents were increased by 1.85-3.82-fold. In the case of Rg1 ginsenoside after MJ elicitation, the content was affected negatively in ginseng hairy root cultures.

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“…Up regulation or down regulation of phytosterol and triterpene production by genetic engineering of Panax species is an attractive strategy to achieve a higher medicinal value (Yue and Zhong 2005). A more detailed understanding of genes involved in saponin biosynthesis could facilitate the genetic modification of plants with altered or novel saponin content (Nelson 2006).…”

Section: Introductionmentioning

confidence: 99%

Regulation and differential expression of protopanaxadiol synthase in Asian and American ginseng ginsenoside biosynthesis by RNA interferences

2013

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Asian ginseng (Panax ginseng) and American ginseng (Panax quinquefolium), are thought to be representative plant of Panax species, have important commercial value and are used in worldwide. Panax species produces triterpene saponins called ginsenosides, which are classified into two groups by the skeleton of aglycones, namely dammarane-type and oleanane-type. Dammaranetype ginsenosides dominate over oleanane-type not only in amount but also in structural varieties. Researches shows that the saponins content in American ginseng is higher than that in Asian ginseng, the higher part of ginsenosides is from dammarane-type biosynthesis. It has been proposed that protopanaxadiol derived from dammarenediol-II, is a key hydroxylation by cytochrome P450 for the biosynthesis of ginsenosides, and the gene number of protopanaxadiol synthase has been published independent in Asian ginseng (PgCYP716A47). However, little is known about genes involved in hydroxylation and glycosylation in American ginseng ginsenoside biosynthesis. Here, we first cloned and identified a P450 gene named PqD12H encoding enzymes catalyzed dammarenediol-II to protopanaxadiol by RT-PCR using degenerate primers designed based on sequence homology. In vitro, the ectopic expression of PqD12H in recombinant WAT21 yeast resulted in protopanaxadiol production after dammarenediol-II was added to the culture medium. In vivo, we established both PgCYP716A47 and PqD12H RNAi transgenic. The RT-PCR and HPLC analysis of the final products of protopanaxadiol and protopanaxatriol showed a result that declined level of protopanaxadiol-type and protopanaxatriol-type ginsenosides. It suggested that the P450 synthase content or expression in American ginseng exceed than in Asian ginseng. The result elucidated the evolution relationship of P450s and the reason of different saponins content among Panax species.

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Purification and characterization of UDPG:ginsenoside Rd glucosyltransferase from suspended cells of Panax notoginseng

Cited by 52 publications

References 32 publications

Biomass and content of ginsenosides and polyacetylenes in American ginseng roots can be increased without affecting the profile of bioactive compounds

Biomass and content of ginsenosides and polyacetylenes in American ginseng roots can be increased without affecting the profile of bioactive compounds

Upregulation of ginsenoside and gene expression related to triterpene biosynthesis in ginseng hairy root cultures elicited by methyl jasmonate

Regulation and differential expression of protopanaxadiol synthase in Asian and American ginseng ginsenoside biosynthesis by RNA interferences

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