Enhancing the biosynthesis of salidroside by biotransformation of p-tyrosol in callus culture of Rhodiola rosea L (original) (raw)
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Acta Biologica Cracoviensia Series Botanica, 2000
The study examined tyrosol glucosyltransferase activity and the efficiency of salidroside production in natural and transformed root cultures of Rhodiola kirilowii (Regel) Regel et Maximowicz. Neither enzyme activity nor salidroside accumulation were detected in natural and transformed root cultures maintained in media without tyrosol. To induce TGase activity in biotransformation reactions, tyrosol was added to natural and transformed root cultures on the day of inoculation. The first peak of TGase activity (0.23 U/μg) was detected on day 9 in natural root culture, accompanied by the highest salidroside content (15.79 mg/g d.w.), but TGase activity was highest (0.27 U/μg) on day 15. In transformed root culture, day 18 showed the highest TGase activity (0.15 U/μg), which coincided with the highest salidroside content (2.4 mg/g d.w.). Based on these results, tyrosol was added to the medium on the days of highest previously detected activity of TGase: day 15 for natural root cultures and day 18 for transformed root cultures. This strategy gave significantly higher yields of salidroside than in the cultures supplemented with tyrosol on the day of inoculation. In natural root culture, salidroside production reached 21.89 mg/g d.w., while precursor feeding in transformed root cultures caused a significant increase in salidroside accumulation to 7.55 mg/g d.w. In all treatments, salidroside production was lower in transformed than in natural root cultures.
PLoS ONE, 2013
Tyrosine decarboxylase initializes salidroside biosynthesis. Metabolic characterization of tyrosine decarboxylase gene from Rhodiola crenulata (RcTYDC) revealed that it played an important role in salidroside biosynthesis. Recombinant 53 kDa RcTYDC converted tyrosine into tyramine. RcTYDC gene expression was induced coordinately with the expression of RcUDPGT (the last gene involved in salidroside biosynthesis) in SA/MeJA treatment; the expression of RcTYDC and RcUDPGT was dramatically upregulated by SA, respectively 49 folds and 36 folds compared with control. MeJA also significantly increased the expression of RcTYDC and RcUDPGT in hairy root cultures. The tissue profile of RcTYDC and RcUDPGT was highly similar: highest expression levels found in stems, higher expression levels in leaves than in flowers and roots. The gene expressing levels were consistent with the salidroside accumulation levels. This strongly suggested that RcTYDC played an important role in salidroside biosynthesis in R. crenulata. Finally, RcTYDC was used to engineering salidroside biosynthetic pathway in R. crenulata hairy roots via metabolic engineering strategy of overexpression. All the transgenic lines showed much higher expression levels of RcTYDC than non-transgenic one. The transgenic lines produced tyramine, tyrosol and salidroside at higher levels, which were respectively 3.21-6.84, 1.50-2.19 and 1.27-3.47 folds compared with the corresponding compound in non-transgenic lines. In conclusion, RcTYDC overexpression promoted tyramine biosynthesis that facilitated more metabolic flux flowing toward the downstream pathway and as a result, the intermediate tyrosol was accumulated more that led to the increased production of the end-product salidroside.
2019
Rhodiola imbricata belonging to Crassulaceae family, is a dioecious perennial plant. It is herb indigenous to the TransHimalayan region and exclusively found in Leh-Ladakh valleys of India and border regions of China and Tibet. Various pharmacological activities in R. imbricata viz. hepatoprotective, radioprotective, immunomodulatory etc., are attributed to presence of different phytochemicals such as ptyrosol, salidroside, rosavin and rosin. But the established plant cell cultures of Rhodiola imbricata usually encounter problems of low product yields and high cost, which discourage its commercialization. So, in order to enhance the secondary metabolite content in R. imbricata, we have studied the effect of different abiotic elicitors on marker compound production. Owing to the previous success of different elicitors on other species of Rhodiola, we have performed chemical and physical elicitations on shoot cultures of Rhodiola imbricata. The best shoot growth in elicited cultures w...
Salidroside content in "in vitro" propagated Rhodiola rosea L
The aim of this study was to determinate the quantity of salidroside in one and two year old Rhodiola rosea L. samples, propagated in vitro, in comparison with coeval ones, grown by seeds. The average content of the investigated substance in the rhizomes was 0.64% at the first and 0.61% at the second year and exceeds that of the marker individuals. These results indicate the in vitro propagation as a possible way to receive a plant material of good quality.
Plant Cell Reports, 2007
Salidroside is a novel effective adaptogenic drug extracted from the medicinal plant Rhodiola sachalinensis A. Bor. Because this plant is a rare resource and has low yield, there is great interest in enhancing the production of salidroside. In this study, a putative UDPglucosyltransferase (UGT) cDNA, UGT73B6, was isolated from Rhodiola sachalinensis using a rapid amplification of cDNA ends (RACE) method. The cDNA was 1,598 bp in length encoding 480 deduced amino acid residues with a conserved UDP-glucose-binding domain (PSPG box). Southern blot analysis of genomic DNA indicated that UGT73B6 existed as a single copy gene in the R. sachalinensis genome. Northern blot analysis revealed that transcripts of UGT73B6 were present in roots, calli and stems, but not in leaves. The UGT73B6 under 35S promoter with double-enhancer sequences from CaMV-W and TMV-W fragments was transferred into R. sachalinensis via Agrobacterium tumefaciens. PCR, PCR-Southern and Southern blot analyses confirmed that the UGT73B6 gene had been integrated into the genome of transgenic calli and plants. Northern blot analysis revealed that the UGT73B6 gene had been expressed at the transcriptional level. High performance liquid chromatography (HPLC) analysis indicated that the overexpression of the UGT73B6 gene resulted in an evident increase of salidroside content. These data suggest that the cloned UGT73B6 can regulate the conversion of tyrosol aglycon to salidroside in R. sachalinensis. This is the first cloned glucosyltransferase gene involved in salidroside biosynthesis. Keywords Rhodiola sachalinensis Á Salidroside Á Tyrosol Á UGT Á Overexpression Abbreviations UGT Uridine diphosphate dependent glycosyltransferase 6-BA N6-Benzyladenine NAA a-Naphthalene acetic acid 2,4-D 2,4-Dichlorophenoxyacetic acid Communicated by L. Jouanin.
Proceedings of the Bulgarian Academy of Sciences
Rhodiola rosea L. is a high-altitude medicinal plant with various health-beneficial effects, associated mainly with the phenolic substances salidroside and rosavins (rosavin, rosin and rosarin). The intensive collection of R. rosea rhizomes and roots to be used in folk medicine and pharmacy has led to a decrease of the species natural resources. New opportunities for species conservation and accelerated synthesis of biologically active substances have been provided by various systems for in vitro and ex vitro cultivation. In this study, the salidroside and rosavins content was determined using HPLC analysis in in vitro cultures of R. rosea and further monitored in adapted regenerants grown ex vitro. The targeted secondary metabolites were not detected in the calli and in the roots of 1-month-old regenerants obtained on different nutrient media. The roots/rhizomes of adapted regenerants grown ex vitro in a greenhouse for 1 year and in the mountain for 1, 2 and 3 years produced increa...
Enhanced Biotransformation Capacity of Rhodiola rosea Callus Cultures for Glycosid Production
Plant Cell, Tissue and Organ Culture, 2005
Rhodiola rosea is a promising medicinal plant that produces various glycosides. Recently we developed a successful method for cultivating it in liquid cultures of compact callus aggregates. In a previous study we reported the successful production of the glycosides of R. rosea by biotransformation of cinnamyl alcohol and tyrosol. In the present study we investigated the possibility of further increasing the yields of the biotransformation products by addition of glucose to the culture medium aside from sucrose, which was used earlier as carbon source. Surprisingly, glucose addition doubled the yield of cinnamyl alcohol glycosides. Rosavin was not produced at all when only sucrose was used. When glucose was added the accumulation dynamics of rosin and a recently described derivative glycoside (designed as compound 321) were similar. Both increased during the first days and then remained constant, while other glycoside compounds increased continuously throughout the cultivation. Rosavin reached its maximum concentration after nine days. In contrast to the beneficial effect on cinnamyl alcohol related glycosides the addition of glucose did not affect the accumulation of the tyrosol derivative salidroside.
Salidroside content in Rhodiola rosea L., dynamics and variability
Th e aim of this study was to investigate the dynamics of the salidroside content in the underground parts of Rhodiola rosea L. during two consecutive vegetation periods and its variability. Th e obtained annual trends of the salidroside content expressed similar pattern for both years and for both kinds of the investigated matherials (roots and rhizomes) of each sex. Th e signiffi cant infl uence on the content of salidroside exert the sex, plant part and season.
Study of Biotransformation Compounds in Callusar Culture of Rhodiola Rosea Specie
2015
Rhodiola rosea L. is a well known species of plants, which has been used medicinally for decades, but study of its pharmacological effects and the compounds responsible for it use still continues. We present data about induced accumulation of secondary metabolites and as well the results of biotransformation of cinamic alcohol in callus culture of R. rosea of Carpathian origin under the influence of same stress factors. The presence of secondary metabolites was investigated by HPLC-MS analysis. The obtained results can be used for selection of valuable genotypes and their future cultivation in artificial conditions.
Safety of a Sustainably Produced, Bioengineered, Nature-Identical Salidroside Compound
Nutrients
Bioactive phytochemicals such as salidroside have been studied to understand the beneficial effects of Rhodiola rosea, an herbaceous plant used in traditional medicine to increase energy and treat a variety of health issues. However, Rhodiola plants are often slow-growing, and many are endangered in their native habitats. Thus, there is a need for safe, alternative supplies of key phytochemicals from Rhodiola. The salidroside subject of this safety study is a synthetic biology product from fermentation of a bioengineered E. coli that produces salidroside. Here, we present comprehensive test results that support the safety of salidroside manufactured via a patented sustainable bioengineering manufacturing process. In vitro bacterial reverse mutation assays with the bioengineered salidroside show no mutagenicity in any of the concentrations tested. In vivo toxicity studies in rats show no adverse effects from the salidroside product. Based on the results of these studies, we conclude ...