Mixture Of Glycosides Research Articles

Histochemical and ultrastructural localization of triterpene saponins in Medicago truncatula.

In the Medicago genus, saponins are complex mixtures of triterpene pentacyclic glycosides extensively studied for their different and economically relevant biological and pharmaceutical properties. This research is aimed at determining for the first time the tissue and cellular localization of triterpene saponins in vegetative organs of Medicago truncatula, a model plant species for legumes, by histochemistry and transmission electron microscopy. The results showed that saponins are present mainly in the palisade mesophyll layer of leaves, whereas in stems they are mostly located in the primary phloem and the subepidermal cells of cortical parenchyma. In root tissue, saponins occur in the secondary phloem region. Transmission electron microscopy revealed prominent saponin accumulation within the leaf and stem chloroplasts, while in the roots the saponins are found in the vesicular structures. Our results demonstrate the feasibility of using histochemistry and transmission electron microscopy to localize M. truncatula saponins at tissue and cellular levels and provide important information for further studies on biosynthesis and regulation of valuable bioactive saponins on agronomic relevant Medicago spp., such as alfalfa (Medicago sativa L.). RESEARCH HIGHLIGHTS: The Medicago genus represents a valuable rich source of saponins, one of the most interesting groups of secondary plant metabolites, which possess relevant biological and pharmacological properties. Plant tissue and cellular localization of saponins is of great importance to better understand their biological functions, biosynthetic pathway, and regulatory mechanisms. We elucidate the localization of saponins in Medicago truncatula with histochemical and transmission electron microscopy studies.

THE EFFECT OF POLYDEXTROSE ON RHEOLOGICAL CHARACTERISTICS AND SENSORY PROPERTIES OF REDUCED SUCROSE STARCH-MILK DESSERTS

Introduction. The aim of the study was to assess the impact of replacing sucrose with polydextrose (E 1200) on the rheological characteristics and sensory properties of starch-milk desserts sweetened with steviol glycosides (E 960). The study materials were desserts made of oxidized potato starch (E 1404), skimmed milk powder, curcumin, vanilla flavorsucrose and/or a mixture of polydextrose (E 1200) and steviol glycosides (E 960). As part of the rheological analyses, the pasting characteristics, viscosity curves and mechanical spectra of the desserts were determined. As part of the sensory analysis, palatability and texture were assessed using a quantitative descriptive analysis. The acceptability of the desserts was also assessed. Results and discussion. Based on the results, it was found that the partial or complete elimination of sucrose from the composition of desserts and replacing it with a mixture of polydextrose and steviol glycosides led to a reduction in their maximum viscosity determined at the heating stage in the pasting test, but did not change the starch pasting temperature. The desserts showed shear-thinning non-Newtonian fluid properties, and the modification of the recipe resulted in a reduction of the consistency coefficient and a reduction in the share of elastic properties without a significant impact on the viscous properties of the system. The desserts in which sucrose was completely, or in a greater degree, replaced with polydextrose and steviol glycosides had a cooling effect and had a licorice aftertaste in the sensory assessment, while retaining sweetness similar to the sweetness of the dessert sweetened with sucrose only. The modification of the dessert recipe did not affect the adhesiveness and mouth melting of the finished products, but contributed to a reduction in their viscosity and malleability assessed by the sensory analysis. Both the texture and palatability of the desserts prepared to the recipe, in which two-third of the amount of sucrose or its total amount was replaced with polydextrose and steviol glycosides, turned out to be less sensorily acceptable than the dessert which did not contain polydextrose.

New HPLC pattern-oriented approach for quality control of enteric coated tablets containing aescin from Aesculus hippocastanum

From the seed of Aesculus hippocastanum L., a complex mixture of triterpene saponin glycosides known as aescin was isolated, purified and characterized to be an active pharmaceutical ingredient for preparing of drug products. A reversed phase high performance liquid chromatographic method with UV detection was developed for the determination of total triterpene glycosides in enteric coated tablets containing aescin in a pattern-oriented manner. The mobile phase consisted of a mixture of acetonitrile and 0.1 % phosphoric acid solution (40: 60). UV detection was performed at 220 nm. The chromatographic column was Gemini C18 column, 5 µm 250 mm × 4.6 mm, which was maintained at 25 ◦C, connected with a precolumn (C18, 4.0 × 3.0 mm, 5 µm). The linear range was from 53.4 to 160.1 μg·mL−1. The RSD of intra-and inter-day precision variations were less than 1 % and the mean recovery was (100.66 ± 0.49) % (RSD = 0.64 %). The method showed its suitability, simplicity, rapidity and precision. Accordingly, it was proven adequate for quality control of this herbal drug product either for registration with the regulartory authority or routine analysis.

Investigation of non-Saccharomyces yeasts with intracellular β-glycosidase activity for wine aroma modification.

Since high proportions of aroma-relevant molecules in plant-derived juices are present in glycosylated forms, the introduction of glycosidase activity during processing is an important tool to modify the aroma composition of the product. During winemaking, the addition of β-glycosidase enzyme or microorganisms with β-glycosidase activity is an established technology. However, low stability under acidic conditions and low selectivity for hydrolysis of different glycosides are still drawbacks, which limit application possibilities. Here, we report the identification and characterization of non-Saccharomyces yeast strains with relatively high β-glycosidase activity in their cultures. We found strong indications for intracellular localization of the enzymes, which is in line with the pH robustness found in experiments with whole cells. Furthermore, we compared the selectivity of aroma compound release from glycoside mixtures using whole cells or cell extracts. The results showed strong differences for the released aroma patterns, which indicates the transport of glycosides and intracellular hydrolysis. Our work demonstrates the application potential of yeasts with intracellular β-glycosidase activities as catalysts with high pH robustness and selective aroma release properties. PRACTICAL APPLICATION: The yeast strains identified and characterized within this work can be applied in wine processing but also in other processes to release aroma molecules from their glycosylated precursors provided by the plants. The strains show relatively high activity of the relevant enzyme, β-glycosidase, also at low pH, which is essential in many processes. In contrast to most other approaches, the enzyme is inside the cells, which can lead to a specific release of certain aroma compounds.

FC090: Stevia as an Alternative Osmotic Agent for Peritoneal Dialysis Fluid

Abstract BACKGROUND AND AIMS Peritoneal dialysis (PD) is a renal replacement technique that requires repeated contact of the peritoneum with hyperosmolar fluids. Unfortunately, it generates metabolic and biomechanical insults that promote structural and functional alterations of the peritoneal membrane (PM). PM is lined by a mesothelial cells (MCs) monolayer that is in direct contact with the PD fluids (PDFs). Repeated PDF exposure causes a mesothelial–mesenchymal transition (MMT) of the MCs, as well as an inflammatory response, angiogenesis and fibrosis of the peritoneal tissue. For decades, glucose (Gl) has been the most commonly used osmotic agent, and Gl itself is responsible for the aforementioned changes. Furthermore, Gl degradation products (GDPs) contribute to deleterious effects on the PM [1]. Therefore, there is an unmet need for more biocompatible osmotic agents able to better maintain the structure and function of the PM. Here we evaluated the biocompatibility of Stevia (STV), a compound of natural origin with a mixture of glycosides. STV has a molecular weight close to the theoretical ideal weight for an osmotic agent in PD [2] and it was used instead of Gl for PDF. METHODS The osmotic capacity of STV-containing fluids was analysed using an in vitro device with artificial membranes and compared to Gl-based fluids. In order to investigate the STV biocompatibility, human omental peritoneal mesothelial cells (HOMCs) were cultured in the presence of different types of Gl-based PDFs (Conventional 4.25% Gl solution with a high GDP level and biocompatible 2.3% Gl solution with low GDPs level), STV-based fluids or TGF-b for 48 h. Mice submitted to surgery of an intraperitoneal catheter insertion were treated for 40 days with STV- or Gl-based PDFs. Peritoneal tissues were collected to determine thickness, MMT, angiogenesis and inflammatory markers, as well as effluents to analyse infiltrating cells. RESULTS Experimentally, using artificial membranes we demonstrated that 1, 0.75 and 0.5% STV-based fluids have a maintained and prolonged osmotic capacity similar to commercial PDFs containing Gl concentrations between 1.5 and 2.3%. Moreover, 1.5% STV-based fluid showed an osmotic capacity higher than a PDF with Gl 2.3% and reached values similar to Gl 4.25% at longer time points. In addition, HOMCs exposed to STV-based fluids showed higher viability and did not induce MMT. Gl-based fluids induced upregulation of mesenchymal markers (VEGF, fibronectin, snail) and also downregulation of the epithelial marker E-cadherin in the case of the highest concentration of Gl (4.25%). None of these markers were significantly affected by STV treatment. In vivo, STV-based fluids preserved MCs monolayer and induced less PM thickness (Figure 1), angiogenesis, leukocyte infiltration and MMT than Gl-based fluids. Furthermore, STV solutions did not promote the release of pro-inflammatory cytokines by HOMCs nor by cells present in mice parietal peritoneal tissue in contrast to Gl-based PDFs. CONCLUSIONS STV-based fluids showed better biocompatibility than Gl-based PDFs in vivo and ex vivo since STV did not induce PM thickness, MMT, angiogenesis or the release of pro-inflammatory cytokines.

FORMULASI DAN UJI EKTIVITAS ANTIBAKTERI SEDIAAN LOTION EKSTRAK ETANOL DAUN SAWAO TERHADAP BAKTERI Stapylococcuc aureus dan Escherchia coli

Manila sapodilla leaves contain phytochemical mixtures of alkaloids, flavonoids, saponins, tannins, terpenoids and glycosides which are known to have antibacterial movement of Staphylococcus aureus and Escherichia coli. The reason for this study was whether the planning of sapodilla leaf extricate salve can be utilized as an enemy of bacterial technique. This exploration was led utilizing a trial which plans to decide all side effects or impacts that emerge, because of specific medicines. Sawo Leaf Ethanol Extract was completed by submerging maceration utilizing 96% ethanol. Antibacterial action testing was done on Sawo leaf extricate with different centralizations of 5%, 10% and 20% and Chaladine cream as a positive control was completed utilizing the discdiffusion strategy. The consequences of the restraint zone movement of Staphylococcus aureus antibacterial lotio arrangements Chaladine sapodilla leaf remove 18 mm, Extract focus 5% 14.5 mm, Extract fixation 10% 20.3 mm, Extract focus 20% 15.46 mm, The aftereffects of the inhibitory zone action in the cream readiness antibacterial concentrate of sapodilla lotio leaf Escherichia coli lotio Chaladine 14 mm, Extract focus 5% 14.16 mm, Extract fixation 10% 20.3 mm, Extract fixation 20% 16.06 mm > 0.05, this demonstrates that the information are typically dispersed, homogeneity trial of 2 gatherings of microscopic organisms and shows a meaning of p > 0.05, showing that the information utilized came from homogeneous information. This demonstrates that Lotio ethanol concentrate of sapodilla leaves has antibacterial movement.

A UV/Vis spectrophotometric methodology for quality control of stevia-based extracts in the food industry

This study established a methodology for quality control of stevia extracts in food industry where one, or a mixture of major stevia glycosides, rebausioside A and stevioside, are present. The methodology relies on pre-treatment to eliminate glucose that may be already present, prior to acid hydrolysis of stevia glycosides toward isosteviol plus glucose and ensuing released glucose quantification using the glucose oxidase/peroxidase coupled assay. Subsequent isosteviol isolation and concentration assessment using UV spectroscopy, permits the quantification of an average number of glucose molecules released per a generated molecule of isosteviol. The latter increases the assay specificity, allowing confirmatory assessment of stevia glycoside identity and purity. Favorable technical characteristics, satisfying international specifications for method validation, were determined for each individual assay (glycoside quantification by acid hydrolysis and isosteviol quantification) for rebaudioside A and stevioside: Standard curves were linear (R2 ≥ 0.993 [R2 ≥ 0.984 for the glucose/isosteviol quantification assay]) over a concentration range from approximately 10−1 mg/mL for the acid assay and 3.0 mg/mL for the glucose/isosteviol assay up to at least 10 mg/mL glycoside. The CV% on all quality control levels was ≤6.33% for repeatability and ≤12.03% for intermediate precision. Bias was ≤±5.00% in all cases. Upon comparison with the gold standard LC/UV method for rebA quantification, we demonstrated a very good agreement and acceptable bias (slope: 1.02, y-intercept: −0.73, R2: 0.994). Acid hydrolysis was tested successfully on commercial products, stevia extracts and commodity sweeteners, containing either of the two major glycosides. We demonstrated that the proposed methodology was able to distinguish between commercial samples containing pure rebaudioside A or stevioside alone and samples containing additional steviol glycosides in combination. When verifying glycoside content in binary mixtures of rebaudioside A and stevioside, bias was sufficiently low (<10%). The proposed methodology requires minimal equipment and expertise and is applicable to a significant percentage (approximately 50%) of all commercial stevia extracts and sweeteners. Our findings should provide a useful tool for stevia quality control, thus allowing an increase in the frequency and extent of testing, even in minimally equipped settings, while containing cost.

Unfolding the apoptotic mechanism of antioxidant enriched-leaves of Tabebuia pallida (lindl.) miers in EAC cells and mouse model

Ethnopharmacological relevanceTabebuia pallida (Lindl.) Miers (T. pallida) is a well-known native Caribbean medicinal plant. The leaves and barks of T. pallida are used as traditional medicine in the form of herbal or medicinal tea to manage cancer, fever, and pain. Moreover, extracts from the leaves of T. pallida showed anticancer activity. However, the chemical profile and mechanism of anticancer activity of T. pallida leaves (TPL), stem bark (TPSB), root bark (TPRB) and flowers (TPF) remain unexplored. Aim of the studyThe present study was designed to explore the regulation of apoptosis by T. pallida using Ehrlich Ascites Carcinoma (EAC) cultured cells and an EAC mouse model. LC-ESI-MS/MS was used for compositional analysis of T. pallida extracts. Materials and methodsDried and powdered TPL, TPSB, TPRB and TPF were extracted with 80% methanol. Using cultured EAC cells and EAC-bearing mice with and without these extracts, anticancer activities were studied by assessing cytotoxicity and tumor cell growth inhibition, changes in life span of mice, and hematological and biochemical parameters. Apoptosis was analyzed by microscopy and expression of selected apoptosis-related genes (Bcl-2, Bcl-xL, NFκ-B, PARP-1, p53, Bax, caspase-3 and -8) using RT-PCR. LC-ESI-MS analysis was performed to identify the major compounds from active extracts. Computer aided analyses was undertaken to sort out the best-fit phytoconstituent of total ten isolated compounds of this plant for antioxidant and anticancer activity. ResultsIn EAC mice compared with untreated controls, the TPL extract exhibited the highest cancer cell toxicity with significant tumor cell growth inhibition (p < 0.001), reduced ascites by body weight (p < 0.01), increased the life span (p < 0.001), normalized blood parameters (RBC/WBC counts), and increased the levels of superoxide dismutase and catalase. TPL-treated EAC cells showed increased apoptotic characteristics of membrane blebbing, chromatin condensation and nuclear fragmentation, and caspase-3 activation, compared with untreated EAC cells. Moreover, annexin V-FITC and propidium iodide signals were greatly enhanced in response to TPL treatment, indicating apoptosis induction. Pro- and anti-apoptotic signaling after TPL treatment demonstrated up-regulated p53, Bax and PARP-1, and down-regulated NFκ-B, Bcl-2 and Bcl-xL expression, suggesting that TPL shifts the balance of pro- and anti-apoptotic genes towards cell death. LC-ESI-MS data of TPL showed a mixture of glycosides, lapachol, and quercetin antioxidant and its derivatives that were significantly linked to cancer cell targets. The compound, pelargonidin-3-O-glucoside was found to be most effective in computer aided models. ConclusionsIn conclusion, the TPL extract of T. pallida possesses significant anticancer activity. The tumor suppressive mechanism is due to apoptosis induced by activation of antioxidant enzymes and caspases and mediated by a change in the balance of pro- and anti-apoptotic genes that promotes cell death.

Morphological characterization of Colchicine-induced Mutants in Stevia rebaudiana

Stevia rebaudiana is a sweet herb that belongs to the family Asteraceae. Steviol glycosides, a natural sweetener derived from Stevia leaves are 100–300 times sweeter than sucrose and contain a complex mixture of sweet diterpene glycosides. The mutagenic effect of colchicine to improve leaf and steviol glycosides yield in stevia was investigated. Stevia plant were treated with five different colchicine concentrations (0.5%, 1.00%, 1.50%, 2.0%, 2.5% and untreated plantlet (0.0%) act as control) for 48h. The plants were arranged in a completely randomized block design. Treated plants were characterized based on morphological traits such as plant height, leaf length and leaf thickness. The results obtained revealed significant difference (p ≤ 0.05) in the morphological traits of the mutants when compared with the control. Overall, 2.0% colchicine concentration resulted in higher average plant height by 36.0% ± 0.6 leaf length increments by 43.0%± 0.7 and the leaf thickness increment by 5.0%± 0.1. The improvement of the mutants’ traits is concentration dependent and increases with increment in colchicine concentration. Thus, we therefore suggested that 2.0% concentration should be employed in improving stevia growth and yield related traits.

Anti-fatigue activity of gardenia yellow pigment and Cistanche phenylethanol glycosides mixture in hypoxia

The plateau, which has a unique geographical environment, forms extremely harsh climate conditions. Compared with that of people in the plains, the athletic ability of people on the plateau is significantly decreased. People who enter the plateau from the plains for short time periods will suffer from metabolic dysfunction, productivity loss, bradykinesia and a significant decrease in overall exercise endurance, which results in exercise fatigue. To investigate the anti-fatigue activity of gardenia yellow pigment and Cistanche phenylethanoid glycosides mixture under hypoxia, we performed anti-hypoxia, normoxia anti-fatigue and hypoxia anti-fatigue experiments. The gardenia yellow pigment and Cistanche phenylethanoid glycosides mixture has good anti-hypoxia and anti-fatigue effects under hypoxia, can significantly prolong the anti-anoxia time of mice, and prolong the exhausted swimming time of mice under normoxic and hypoxic conditions. The mechanisms may be related to reducing the accumulation of bad metabolites, increasing energy reserves, improving free radical scavenging capacity, increasing related metabolic enzymes activities, reducing apoptosis and inhibiting the infiltration of inflammatory cells. The gardenia yellow pigment and Cistanche phenylethanoid glycosides mixture is a potential functional product to improve hypoxic exercise fatigue.

Antioxidant and anti-diabetic potential of rebaudioside A and a mixture of steviol glycosides in alloxan-induced diabetic rats

Diabetes mellitus, a metabolic disorder rapidly spreading worldwide affects multiple organs and body functions. Stevia rebaudiana (Bertoni), belonging to the Asteraceae family is a herb with medicinal and commercial importance to cure chronic diseases like diabetes mellitus. The present study was aimed to explore the antidiabetic effect of two reference standards like Steviol Glycosides System Suitability (SGSS, a mixture of nine steviol glycosides) and Rebaudioside A in alloxan-induced diabetic rats. In this regard, diabetes was induced in rats by administration of a single intraperitoneal (i.p.) injection of alloxan monohydrate (150 mg/kg body weight). After exposure to alloxan monohydrate, the overall intracellular antioxidant functions get disturbed which significantly increase the blood glucose levels, plasma malondialdehyde and sialic acid content. The oral administration of SGSS and Reb A at a dose of (20 and 30 mg/kg b.wt.) for a period of 21 days to diabetic rats significantly (P ˂0.05) reduced the blood glucose levels when compared with alloxan-induced diabetic control (DC) rats of group II and restore the antioxidant potential by decreasing the oxidative damage and also normalizes hyperlipidemic indices. The study demonstrated that bioactive components (steviol glycosides) of S. rebaudiana have potential therapeutic prospects to attenuate diabetes and its related complications.

Chemical study of the stem bark of Phyllanthus emblica (Phyllanthaceae)

AbstractFrom the ethyl acetate extract of the stem bark of Phyllanthus emblica, a mixture of triterpene glycosides (1) and six compounds (2‐7) were isolated. The structure of these compounds were established as 3‐O‐α‐L‐arabinopyranosyloleanolic acid (1a), 3‐O‐α‐L‐arabinopyranosylursolic acid (1b), 16‐dehydropregnenolone (2), periplogenin (3), betulin (4), phyllanthol (5), gallic acid (6) and methyl gallate (7) by 1D, 2D NMR spectroscopies, and high resolution ESI‐MS analysis, as well as comparison with those reported in the literature. The compounds (1a), (1b), (2) and (3) were found for the first time from this genus.

Metabolic fate in adult and pediatric population of steviol glycosides produced from stevia leaf extract by different production technologies

More than 60 naturally occurring steviol glycosides in the Stevia rebaudiana Bertoni plant share a similar molecular structure with an aglycone steviol backbone conjugated with β- and α-glycosidic bonds to different sugar moieties. These glycosides are naturally produced in different quantities within the stevia leaf. Certain minor glycosides with superior sensory attributes, such as Reb D and Reb M, are found less than 0.1% in traditional stevia leaves. New technologies can now produce better tasting steviol glycosides by using enzymatic conversion of stevioside and Reb A, which are abundant in stevia leaf. Several regulatory authorities recently evaluated steviol glycosides produced by enzymatic conversion of stevia leaf extract and approved them safe for human consumption.Steviol glycosides undergo microbial hydrolysis in the colon to generate steviol, which is absorbed and metabolized into steviol glucuronide, and excreted primarily via human's urine. Previous studies have shown the hydrolysis of highly purified individual steviol glycosides extracted from stevia leaf are converted to steviol in the presence of colonic microbiota of adults. Since colonic microbiota of children may be different from adults, this study investigates the metabolic fate in the colonic microbiota of adults and children of the minor steviol glycosides produced by extraction and enzymatic conversion of major steviol glycosides from stevia leaf.Several in vitro incubation tests were conducted in human fecal homogenates collected from adult and pediatric populations with steviol glycoside test samples comprised of a complex stevia leaf extract, a blend of minor glycosides isolated from stevia extract and two mixtures of steviol glycosides produced by enzymatic conversion of Reb A to larger molecules by attaching glucose units via β- or α-glycosidic bonds. Results from these studies clearly demonstrate steviol glycosides produced by extraction from stevia leaf, or enzymatic conversion of stevia leaf extract, share the same metabolic fate in the human gut microbiota from adults and children. Considering a common metabolite structure and a shared metabolic fate in all ages, safety data for individual steviol glycosides can be used to support safety of all steviol glycosides produced by extraction and enzymatic conversion of stevia leaf extract.

Surface properties and microemulsion of anionic/nonionic mixtures based on sulfonate Gemini surfactant in the presence of NaCl

The mixing properties of anionic/nonionic surfactant mixtures in the presence of inorganic salts are essential to the industrial application. The equilibrium and dynamic surface tension of the sulfonate Gemini surfactant (SGS12) and alkyl glycoside (APG0810) mixtures were examined using the plate method and drop-shape analysis. Moreover, the Winsor phase diagram and solubility properties of the microemulsion were studied using the salinity scan. Results show that the critical micelle concentration (cmc) of the SGS12/APG0810 mixtures was intermediate between those of the two single surfactants in the presence of NaCl. For the SGS12/APG0810 mixtures, the absolute value of interaction parameters increased as the APG0810 content increased at cNaCl = 0.01 or 0.1 mol/L. The formation of the mixed micelles was spontaneous. At cNaCl = 0.01 mol/L, the initial adsorption (t → 0) of surfactant molecules from the bulk phase to the surface was diffusion-controlled. The salinity scan experiments showed that the optimal solubilization parameters of the SGS12/APG0810 microemulsion decrease with the decrease of the molar fraction of SGS12. However, the concentration range of inorganic salt between the formation and disappearance of Winsor III broadened with the decline in SGS12 content. The impact of temperature on the volume fraction of the middle-phase microemulsion (MPM) was ignorable, indicating the high stability of MPM at the temperature range of 30.0 °C ~ 80.0 °C. At 30.0 ± 0.1 °C, the hydrodynamic diameter of the SGS12/APG0810 MPM was about 28.0 nm, and the number of microemulsion droplets decreased with the increase of APG0810 content. When the total mass fraction of surfactant and cosurfactant was 15%, wNaCl = 5% (brine), Km = 1:3 and Vo/Vw = 1, the interfacial tension between crude oil and the SGS12/APG0810 MPM reached the order of 10−3mN/m. At nSGS12:nAPG0810 = 7:3, the lowest interfacial tension was 9 × 10−4 mN/m. For the SGS12/APG0810 (7:3) mixture, the addition of n-butanol induced the formation of MPM. The concentration range of n-butanol between the formation and the disappearance of Winsor III first increased and then decreased with the increase of surfactant concentration.

ABCC Transporters Mediate the Vacuolar Accumulation of Crocins in Saffron Stigmas.

Compartmentation is a key strategy enacted by plants for the storage of specialized metabolites. The saffron spice owes its red color to crocins, a complex mixture of apocarotenoid glycosides that accumulate in intracellular vacuoles and reach up to 10% of the spice dry weight. We developed a general approach, based on coexpression analysis, heterologous expression in yeast (Saccharomyces cerevisiae), and in vitro transportomic assays using yeast microsomes and total plant metabolite extracts, for the identification of putative vacuolar metabolite transporters, and we used it to identify Crocus sativus transporters mediating vacuolar crocin accumulation in stigmas. Three transporters, belonging to both the multidrug and toxic compound extrusion and ATP binding cassette C (ABCC) families, were coexpressed with crocins and/or with the gene encoding the first dedicated enzyme in the crocin biosynthetic pathway, CsCCD2. Two of these, belonging to the ABCC family, were able to mediate transport of several crocins when expressed in yeast microsomes. CsABCC4a was selectively expressed in C. sativus stigmas, was predominantly tonoplast localized, transported crocins in vitro in a stereospecific and cooperative way, and was able to enhance crocin accumulation when expressed in Nicotiana benthamiana leaves.plantcell;31/11/2789/FX1F1fx1.

Efficient bioconversion of epimedin C to icariin by a glycosidase from Aspergillus nidulans

Herba Epimedii is a traditional Chinese herbal medicine that contains a mixture of bioactive flavonoid glycosides. Among them, icariin has the most outstanding bioactive functions, while epimedin C exhibits substantial toxicity. A recombinant α-L-rhamnosidase (synAnRhaE) from Aspergillus nidulans was expressed in Escherichia coli to promote the efficient bioconversion of epimedin C to icariin. A hydrolase activity of 574.5 U L−1 was acquired via optimized fed-batch fermentation in a 5-L bioreactor. The enzyme proved to be stable in an acidulous pH range below 55 °C with an optimal pH of 4.5 and optimal temperature of 55 °C. Epimedin C (1 g L−1) was 100% converted to icariin within 90 min using recombinant cells. The resting cells proved to be selective for epimedin C and 2″-O-rhamnosylicariside II in crude extracts of the epimedium plant. This work provides an original and efficient biocatalyst system that can be applied in industrialized production of icariin.

Antioxidant activity, phenol and flavonoid content, and formulation cream of Stevia rebaudiana Bert

The presented study aimed to characterize the content, biological activity and formulation of extracts prepared from dried Stevia rebaudiana Bert leaves. Stevia rebaudiana Bert is the leaves that have antioxidant activity. The leaves, stems and flowers of stevia contain a complex mixture of sweet diterpene glycosides, including isosteviol, stevioside, rebaudioside (A, B, C, D, E, and F), steviolbioside and dulcoside A. In addition to glycosides, the leaves of stevia also contain other phytoconstituents, such as flavonoids, phenolic acids, fatty acids, proteins, and vitamins. These compounds can prevent and retain free radicals substitution, which can cause aging and chronic diseases. The extract obtained was filtered and evaporated — determination of total phenolics and total flavonoid using spectrophotometric methods. The antioxidant potential of this extract was evaluated by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. In the DPPH radical-scavenging activities, the extract had the antioxidant activity (IC50 = 48.18 μg / ml). The results showed that these extracts of Stevia rebaudiana Bert could be considered as natural antioxidants. The evaluation of the formulated cream showed good results and can be good potential for cosmetic product development.

Utilization Of Sterol Glycosides In Fame (Fatty Acid Methyl Ester) Byproducts From The Biodiesel Industry

In recent years, the development of renewable energy such as biodiesel has been widely researched throughout the world as technology advances in the era of Industry 4.0. At the final station of biodiesel production in the maturation tank, the by-products will form by-products in the form of sterol glycosides in Fatty Acid Methyl Esters which have not been utilized. This study aims to determine the volume of biodiesel from a mixture of sterol glycosides with a ratio of 0.5% H2SO4 catalyst concentration; 1%; 1.5% and 2% and tested their characteristics in accordance with the Indonesian National Standard (SNI 7182: 2015). Biodiesel production was carried out by esterification with a molar ratio of 1:6 (sterol glycoside: methanol) to variations in H2SO4 catalyst concentration. The results of the highest yield volume biodiesel were obtained from a catalyst concentration of 1.5% of 28.02% and the lowest yield of 17.50% in a 0.5% catalyst. Based on the characteristic test of biodiesel by varying the concentration of catalyst H2SO4 obtained density of 852 – 862 kg m-3, viscosity of 4.642 – 4.950 mm2 s-1 and saponification number of 191.007 – 198.164 mg-KOH g-1 according to standard characteristics SNI 7182:2015, while for the water content of 0.1965 – 0.1976% and acid numbers of 2.151 – 3.232 mg-KOH g-1 isn’t according to standard characteristics. Based on research, pre-treatment treatments was recommended before the refining process to reduce the amount of acid and moisture content so according to standard characteristics.

The Regulatory Effects of Paeoniflorin and Its Derivative Paeoniflorin-6'-O-Benzene Sulfonate CP-25 on Inflammation and Immune Diseases.

The plant extract “total glucosides of peony” (TGP) constitutes a mixture of glycosides that is isolated from the roots of the well-known traditional Chinese herb Paeonia lactiflora Pall. Paeoniflorin (Pae) is the most abundant component and the main biologically active ingredient of TGP. Pharmacologically, Pae exhibits powerful anti-inflammatory and immune regulatory effects in some animal models of autoimmune diseases including Rheumatoid Arthritis (RA) and Systemic Lupus Erythematosus (SLE). Recently, we modified Pae with an addition of benzene sulfonate to achieve better bioavailability and higher anti-inflammatory immune regulatory effects. This review summarizes the pharmacological activities of Pae and the novel anti-inflammatory and immunomodulatory agent Paeoniflorin-6′-O-benzenesulfonate (CP-25) in various chronic inflammatory and autoimmune disorders. The regulatory effects of Pae and CP-25 make them promising agents for other related diseases, which require extensive investigation in the future.

Bioassay-guided isolation of cytotoxic compounds from Chrysophthalmum montanum (DC.) Boiss

Bioassay-guided isolation of the 80% methanol extract of the aerial parts of Chrysophthalmum montanum (DC.) Boiss. (Asteraceae) led to the isolation of four known guaianolide-type sesquiterpene lactones, 6α-acetoxy-4α-hydroxy-1βH-guaia-9.11(13)-dien-12.8α-olide (1), 6α-acetoxy-4α-hydroxy-9β.10β-epoxy-1βH-guaia-11(13)-en-12.8α-olide (2), 4α,6α-dihydroxy-1β,5α,7αH-guaia-9(10),11(13)-dien-12,8α-olide (3), and (4α,5α,8β,10β)-4,10-dihydroxy-1,11(13)-guaidien-12,8-olide (4), along a steroidal glycoside mixture (5a and 5b). The structures of the compounds were identified on the basis of spectroscopic data. Among them, 2, 4 and a steroidal glycoside mixture were obtained from C. montanum for the first time. All isolates were also first time assayed for in vitro cytotoxicities against four human cancer cell lines, i.e. breast (MCF-7, MDA-MB 231), colon (HT-29), and lung (PC3). Among the isolates, 1–3 showed significant inhibitory effect on the proliferation of cancer cells with viability ranging from 6.86 to 26.51%, while steroidal glycoside mixture showed no cytotoxicity, except against HT-29 (viability 61.99%). Compound 4 exhibited strong and selective cell growth inhibition against HT-29 with viability 20.99% and was identified as a promising compound with high selectivity between cancer cells and normal human lung cells (BEAS-2B), especially against HT-29 (IC50 = 12.2 μg/mL) compared to that of cisplatin. These results suggested that 4 is worthy of further study to determine its cytotoxicity mechanisms.