Plant Glycosides Research Articles

The Role of Plant Extracts in Enhancing Nutrition and Health for Dogs and Cats: Safety, Benefits, and Applications.

Plant extracts, derived from various natural sources, encompass primary and secondary metabolites, which include plant polysaccharides, polyphenols, alkaloids, flavonoids, glycosides, terpenes, and volatile oils. These compounds exhibit a range of biological activities such as antioxidant, anti-inflammatory, and antimicrobial functions. Currently, polyphenols and other bioactive compounds are being incorporated into the diets of farm animals, fish, and pets to promote health benefits. Despite this, the application and potential of plant extracts in canine and feline nutrition have not been comprehensively explored. Many aspects of the mechanisms underlying the action of these plant metabolites remain to be analyzed and elucidated. Furthermore, leveraging natural plant extracts for the treatment of clinical conditions in dogs and cats is a crucial component of clinical nutrition. Consequently, this review aims to highlight the impact of plant extracts on overall health, gastrointestinal health, immune health, cardiovascular health, redox balance, and pathology in dogs and cats.

Plant P450 forms indigo and indirubin when expressed in Escherichia coli

Indigo and indirubin are derived from indoxyl molecules, which generally occur as indoxyl glycosides in woad (Isatis tinctoria L.) and other indigo-producing plants. Indoxyl glycosides are biosynthesized from indole via 3-hydroxylation to form indoxyl, followed by one or more glycosylations. Enzymes that attach and remove sugars to and from indoxyl have already been isolated and characterized, while enzymes that convert indole into indoxyl in plants have remained elusive, until the identification of P450s and flavin-containing monooxygenases that hydroxylate indole. A P450 gene from woad (named CYP71B102) was heterologously expressed in E. coli, resulting in the formation of indigo and indirubin, as well as isatin and 2-oxindole, which along with indoxyl are putative precursors of indirubin. The addition of either isatin or 2-oxindole to the recombinant E. coli reduced the levels of indigo and increased the amount of indirubin, whereas coexpression of CYP71B102 with isatin hydroxylase (which degrades isatin) increased the levels of indigo and decreased the amount of indirubin, albeit slightly. The results suggest that CYP71B102 hydroxylates indole at both the 2- and 3- positions to produce 2-oxindole and indoxyl, respectively, and that the coupling of indoxyl with either 2-oxindole or isatin forms indirubin, while dimerization of indoxyl forms indigo. This P450 gene is thus likely involved in the biosynthesis of indirubin in woad, as well as the formation of indigo and its glycosidic precursors, even if other types of enzymes, such as flavin-containing monooxygenases, may be involved in indole hydroxylation in other indigo-producing plants.

Metroxylon sagu Rottb. Fruit Flour as Potential Prebiotics for Selected Probiotics and Phytochemical Profiling of its Methanolic Extract by LC-MS/MS

The plant of Metroxylon sago Rottb. which is found abundantly in Indonesia, Malaysia, Philippines, and Papua New Guinea, is famous as carbohydrate producing plants. The sago fruit has not been yet extensively used, although they are rich in oligosaccharides which is potential as prebiotic. Sago fruit flour was studied as growth substrate for five selected probiotics and phytochemical profile of its methanol extracts were determined. All bacteria grew well up to 24 h on the growth medium containing sago fruit flour as carbohydrate substitute. However, Lacticaseibacillus rhamnosus FNCC 0099 and Lactobacillus acidophilus FNCC 0051 showed viability stability up to 72 h, while the others were slightly decreased. The sago fruit flour contained 20.4 mg/g total sugars, 14.8 mg QE/g flavonoids, and 36.7 mg GAE/g phenolics. After extraction of the fruit flour with 80% methanol using an ultrasonicator at 55 °C at 40 kHz for 40 min, the extract was analysed for its phytochemical profiles using untargeted LC-MS/MS screening with negative ionization mode. Seven compounds categorised in three distinct groups i.e., sugar alcohol, plant glycosides and fatty acids, have been identified as having possible prebiotic activity. These include ((1xi)-1,5-anhydro-2,3,6-tris-O-(carboxymethyl)-1-methyl-4-O-methyl-D-glucitol), quercitrin, (15Z)-9,12,13-trihydroxy-15-octadecenoic acid, corchorifatty acid F, 3,5-dihydroxy-2-(4-hydroxyphenyl)-4-oxo-3,4-dihydro-2H-chromen-7-yl-hexopyranoside, gynocardin, and (4-methylumbelliferone)-b-D-glucopyranoside. As the sago fruit flour has been proved as potential prebiotic in this study, its extensive prebiotics activity, prebiotic index, and stability under digestive condition will be subjected for further study.

Phytochemical screening, antimicrobial, and antioxidant properties of Helianthus annuus and Hyophila involuta: A comparative account

Angiosperms and bryophytes, though distantly related plant groups, have many similar ecological and economic implications, including medicinal value. Therefore, this study aimed to analyse the phenolic and flavonoids composition and antioxidant and antimicrobial activities of Helianthus annuus L. (Angiosperms-Asteraceae) and another plant, a moss (Bryophyta), Hyophila involuta (Hook.) Jaeg., aerial parts (leaves), prepared in four different extracts (methanol, chloroform, distilled water, and petroleum ether). Phytochemical screening was conducted using standard methods of precipitation and colouration reactions. The Folin-Ciocalteu method was employed to determine the total phenol content, while the Aluminium Chloride Colorimetric method was used for flavonoid content determination. The antioxidant activity was measured through two methods: DPPH and NOSA scavenging activity. The phytochemical screening detected the presence and absence of fixed oils and fats, flavonoids, saponins, terpenoids, tannins, polyphenols, carbohydrates, and glycosides in both plants. The antibacterial and antifungal activity of both plants' methanolic extracts was examined against bacterial and fungal pathogens, i.e., Escherichia coli and Bacillus subtilis and fungal strains, i.e., Fusarium oxysporum and Aspergillus niger. The results were compared to a regular antibiotic disc and negative control that served as a methanol solvent. The methanolic extract of H. annuus has higher total phenol and flavonoid content, as well as antioxidant and antimicrobial activity, than H. involuta. Based on these data, it can be concluded that, while H. annuus is more effective than H. involuta, both distantly related plant species have similar phytochemical profiles and should be included equally in future herbal compositions.

Plant Cyanogenic-Derived Metabolites and Herbivore Counter-Defences.

The release of cyanide from cyanogenic precursors is the central core of the plant defences based on the cyanogenesis process. Although cyanide is formed as a coproduct of some metabolic routes, its production is mostly due to the degradation of cyanohydrins originating from cyanogenic glycosides in cyanogenic plants and the 4-OH-ICN route in Brassicaceae. Cyanohydrins are then hydrolysed in a reversible reaction generating cyanide, being both, cyanohydrins and cyanide, toxic compounds with potential defensive properties against pests and pathogens. Based on the production of cyanogenic-derived molecules in response to the damage caused by herbivore infestation, in this review, we compile the actual knowledge of plant cyanogenic events in the plant-pest context. Besides the defensive potential, the mode of action, and the targets of the cyanogenic compounds to combat phytophagous insects and acari, special attention has been paid to arthropod responses and the strategies to overcome the impact of cyanogenesis. Physiological and behavioural adaptations, as well as cyanide detoxification by β-cyanoalanine synthases, rhodaneses, and cyanases are common ways of phytophagous arthropods defences against the cyanide produced by plants. Much experimental work is needed to further understand the complexities and specificities of the defence-counter-defence system to be applied in breeding programs.

Evidence for Reductions in Physical and Chemical Plant Defense Traits in Island Flora.

Reduced defense against large herbivores has been suggested to be part of the "island syndrome" in plants. However, empirical evidence for this pattern is mixed. In this paper, we present two studies that compare putative physical and chemical defense traits from plants on the California Channel Islands and nearby mainland based on sampling of both field and common garden plants. In the first study, we focus on five pairs of woody shrubs from three island and three mainland locations and find evidence for increased leaf area, decreased marginal leaf spines, and decreased concentrations of cyanogenic glycosides in island plants. We observed similar increases in leaf area and decreases in defense traits when comparing island and mainland genotypes grown together in botanic gardens, suggesting that trait differences are not solely driven by abiotic differences between island and mainland sites. In the second study, we conducted a common garden experiment with a perennial herb-Stachys bullata (Lamiaceae)-collected from two island and four mainland locations. Compared to their mainland relatives, island genotypes show highly reduced glandular trichomes and a nearly 100-fold reduction in mono- and sesquiterpene compounds from leaf surfaces. Island genotypes also had significantly higher specific leaf area, somewhat lower rates of gas exchange, and greater aboveground biomass than mainland genotypes across two years of study, potentially reflecting a broader shift in growth habit. Together, our results provide evidence for reduced expression of putative defense traits in island plants, though these results may reflect adaptation to both biotic (i.e., the historical absence of large herbivores) and climatic conditions on islands.

Genome-wide analysis of the mulberry (Morus abla L.) GH9 gene family and the functional characterization of MaGH9B6 during the development of the abscission zone.

Plant glycoside hydrolase family 9 genes (GH9s) are widely distributed in plants and involved in a variety of cellular and physiological processes. In the current study, nine GH9 genes were identified in the mulberry and were divided into two subfamilies based on the phylogenetic analysis. Conserved motifs and gene structure analysis suggested that the evolution of the two subfamilies is relatively conserved and the glycoside hydrolase domain almost occupy the entire coding region of the GH9s gene. Only segmental duplication has played a role in the expansion of gene family. Collinearity analysis showed that mulberry GH9s had the closest relationship with poplar GH9s. MaGH9B1, MaGH9B6, MaGH9B5, and MaGH9B3 were detected to have transcript accumulation in the stalk of easy-to drop mature fruit drop, suggesting that these could play a role in mulberry fruit drop. Multiple cis-acting elements related to plant hormones and abiotic stress responses were found in the mulberry GH9 promoter regions and showed different activities under exogenous abscisic acid (ABA) and 2,4- dichlorophenoxyacetic acid (2,4-D) stresses. We found that the lignin content in the fruit stalk decreased with the formation of the abscission zone (AZ), which could indirectly reflect the formation process of the AZ. These results provide a theoretical basis for further research on the role of GH9s in mulberry abscission.

Fabrication of a bio-based polymer adsorbent and its application for extraction and determination of glycosides from Huangqi Liuyi Decoction

Huangqi Liuyi Decoction, a famous classical Chinese prescription, shows significant curative effect on diabetes and its complications, in which calycosin-7-glucoside, liquiritin and glycyrrhizic acid are the main components that playing these mentioned pharmacological activity, under the synergistic action of various other ingredients in the decoction. However, there are significant differences in the content of active compounds in Chinese medicinal materials, which mainly due to origin, picking seasons, and processing methods. Hence, the accurate content of the glycosides is the prerequisite for ensuring the pharmacological efficacy. Aiming at establishing an efficient extraction and determination method for accurate quantitative analysis of calycosin-7-glucoside, liquiritin and glycyrrhizic acid in Huangqi Liuyi Decoction, an on line solid-phase extraction-high-performance liquid chromatography method was developed, using a homemade bio-based monolithic adsorbent. The bio-based adsorbent was prepared in a stainless steel tube, using bio-monomers of methyleugenol and S-allyl-L-cysteine, which effectively reduced the dependence of the polymer field on non-renewable fossil resources and reduced carbon emissions. Furthermore, the prepared adsorbent owned abundant chemical groups, which can produce interactions of hydrogen bond, dipole-dipole, π-π and hydrophobic force with the target glycosides, thus improving the specific recognition ability of the adsorbent. The experiments were carried out on an LC-3000 HPLC instrument with a six-way valve. Methodology validation indicates that the recovery is in the range of 97.0%-103.4% with the RSD in the range of 1.6%-4.0%, due to the specific selectivity of the bio-based monolithic adsorbent for these three glycosides, and good matrix-removal ability for Huangqi Liuyi decoction. The limit of detection is 0.17, 0.50 and 0.33μg/mL for calycosin-7-glucoside, liquiritin and glycyrrhizic acid, respectively, and the limit of quantitation is 0.50, 1.50 and 1.00μg/mL, respectively, with the linear range of 2-200μg/mL for calycosin-7-glucoside, and 5-500μg/mL for liquiritin and glycyrrhizic acid. The present work provided a simple and efficient method for the extraction and determination of glycosides in complex medicinal plants.

Fine-Scale Characterization of Plant Diterpene Glycosides Using Energy-Resolved Untargeted LC-MS/MS Metabolomics Analysis.

Plant diterpene glycosides are essential for diverse physiological processes. Comprehensive structural characterization proved to be a challenge due to variations in glycosylation patterns, diverse aglycone structures, and the absence of comprehensive reference databases. In this study, a method for fine-scale characterization was proposed based on energy-resolved (ER) untargeted LC-MS/MS metabolomics analysis using steviol glycosides as a demonstration. Energy-dependent fragmentation patterns were unveiled by a series of model compounds. Distinct glycosylation sites were discerned by leveraging varying fragmentation energies for the precursor ions. The sugar moiety linkage at C19OOH (R1) exhibited facile and intact cleavage at low collision energies, while the sugar moiety at C13-OH (R2) demonstrated consecutive cleavage with increasing energy. Aglycone ions exhibited a higher relative intensity at NCE 50, with relative intensities ranging from 95% to 100%. Subsequently, aglycone candidates, R1 sugar composition, and R2 sugar sequence were deduced through ER-MS/MS analysis. The developed method was applied to Stevia rebaudiana leaves. A total of 91 diterpene glycosides were unambiguously identified, including 16 steviol glycosides with novel acetylglycosylation patterns. This method offers a rapid alternative for glycan analysis and the structural differentiation of isomers. The developed method enhances the understanding of diterpene glycosides in plants, providing a reliable tool for the in-depth characterization of complex metabolite profiles.

Arbutin interacts with Vibrio harveyi hemolysin to alleviate damage from associated infection

Vibrio harveyi, an opportunistic pathogen, is a major causative agent of vibriosis in aquatic organisms. Many research conducted both domestically and internationally has revealed the significant multidrug resistance exhibited by V. harveyi. Among its important virulence factors, V. harveyi hemolysin (VHH) plays a crucial role in the infection process of this bacterium within the host. Due to the limitations of traditional antimicrobial drugs and the need to prevent bacterial resistance, alternative approaches are being explored worldwide. In this study, VHH was targeted, and through drug screening, it was discovered that arbutin, a plant glycoside, effectively inhibits the hemolytic activity of the VHH. The findings indicate that arbutin does not impede the growth of V. harveyi in vitro, nor does it affect the transcription of related virulence factors such as vhh and luxR. Instead, arbutin counteracts the hemolytic activity of VHH by directly binding to key amino acid residues, including Tyr227, Asn159, and Trp200, thereby influencing the function of the cytotoxic protein. In the context of V. harveyi infection, the presence of arbutin enhances cell survival in vitro and mitigates pathological damage to the gills and kidneys of experimental animals. The results show that arbutin, has the potential to be a promising anti-virulence therapy.

SUPRAMOLECULAR COMPLEXES OF GLYCYRRHISIC ACID AND ITS MONOAMMONIUM SALT WITH ETHACISIN HYDROCHLORIDE

The problem of effective and safe therapy of cardiac arrhythmias requires the creation of new antiarrhythmic drugs with low toxicity. Clathration of medicinal substances with cyclodextrins or plant glycosides is a promising method for reducing their side effects and increasing their solubility.
 The aim of this research is the complex formation of ethacizin hydrochloride with triterpene glycosides and the study of physicochemical properties of the resulting complexes.
 Complexes of glycyrrhizic acid (GA) and its monoammonium salt (MASGA) with ethacizine hydrochloride (EtHQ) in various molar ratios (2 : 1, 4 : 1, 8 : 1) characterized by certain physicochemical parameters have been obtained. The resulting inclusion compounds were studied by UV and IR spectroscopy. It has been determined that molecular complexes of GA and MASGA with EtHQ are formed through weak intermolecular interactions, such as hydrogen bonding, electrostatic and hydrophobic interactions. Supramolecular complexes of natural glycosides with EtHQ have the same stoichiometric composition, approximately equal to the stability constant, and the negative value of the Gibbs free energy confirms the formation of a molecular complex as a result of autoassociation.
 The obtained experimental data can serve and/or supplement scientific data for the creation of new promising drugs with a broad therapeutic effect, targeted delivery, synergistic effect and low toxicity.

How to awaken a sleeping giant: antagonistic expression of Flowering locus T homologs and elements of the age-related pathway are associated with the flowering transition in Agave tequilana

Key messageAntagonistic expression of Flowering locus T proteins and the ageing pathway via miRNAs and sugar metabolism regulate the initiation of flowering in A. tequilana.Flowering in commercial plantations of Agave tequilana signals that plants are ready to harvest for tequila production. However, time of flowering is often unpredictable and a detailed understanding of the process would be beneficial in the field, for breeding and for the development of future research. This report describes the functional analysis of A. tequilana FLOWERING LOCUS T (FT) genes by heterologous expression in A. thaliana and in situ hybridization in agave plants. The gene structures of the Agave tequilana FT family are also described and putative regulatory promoter elements were identified. Most Agave species have monocarpic, perennial life cycles that can last over 25 years during which plants do not respond to the normal environmental signals which induce flowering, suggesting that the ageing pathway as described in Arabidopsis may play an important role in determining flowering time in these species. Elements of this pathway were analyzed and in silico data is presented that supports the regulation of SQUAMOSA PROMOTER BINDING LIKE proteins (SPL), APETALA2 (AP2) proteins and members of Plant Glycoside Hydrolase Family 32 (PGHF32) by interactions with miRNAs 156, 172 and 164 during the initiation of flowering in A. tequilana.

A GH81-type β-glucan-binding protein enhances colonization by mutualistic fungi in barley

Cell walls are important interfaces of plant-fungal interactions, acting as robust physical and chemical barriers against invaders. Upon fungal colonization, plants deposit phenolics and callose at the sites of fungal penetration to prevent further fungal progression. Alterations in the composition of plant cell walls significantly impact host susceptibility. Furthermore, plants and fungi secrete glycan hydrolases acting on each other's cell walls. These enzymes release various sugar oligomers into the apoplast, some of which activate host immunity via surface receptors. Recent characterization of cell walls from plant-colonizing fungi has emphasized the abundance of β-glucans in different cell wall layers, which makes them suitable targets for recognition. To characterize host components involved in immunity against fungi, we performed a protein pull-down with the biotinylated β-glucan laminarin. Thereby, we identified a plant glycoside hydrolase family 81-type glucan-binding protein (GBP) as a β-glucan interactor. Mutation of GBP1 and its only paralog, GBP2, in barley led to decreased colonization by the beneficial root endophytes Serendipita indica and S.vermifera, as well as the arbuscular mycorrhizal fungus Rhizophagus irregularis. The reduction of colonization was accompanied by enhanced responses at the host cell wall, including an extension of callose-containing cell wall appositions. Moreover, GBP mutation in barley also reduced fungal biomass in roots by the hemibiotrophic pathogen Bipolaris sorokiniana and inhibited the penetration success of the obligate biotrophic leaf pathogen Blumeria hordei. These results indicate that GBP1 is involved in the establishment of symbiotic associations with beneficial fungi-a role that has potentially been appropriated by barley-adapted pathogens.

Waste orange seeds (Citrus sinensis seed) transformation, a viable industrial bio-oil: Oil extraction, physicochemical characterization and vital instrumental analyses studies

The processing of Petroleum-based oil has contributed to environmental challenges such as climate change. Recently, research attention has been shifted to oil extraction from different non-edible seeds as a suitable substitute. Accordingly, this study examines the physiochemical characterization, and the effect of extraction temperature on the yield of waste Citrus sinensis seed [orange seed (OS)] bio oil. Studies on the prevalent functional group, fatty acid methyl ester compositions and thermal oxidative stability were carried out using the Fourier transform infrared spectroscopy (FTIR), gas chromatography-mass spectroscopy (GCMS) and differential scanning calorimetry (DSC), respectively. The oil content of the OS bio oil was 46.26%w/w. The high iodine number (31.47 mg/100 g) and low pour point (4.50 °C) of the OS bio oil showed its high content of unsaturated fatty acid. The FTIR functional group of the OS bio oil showed predominantly alkane of CH stretching, aldehydes, esters, and carboxylic acid. The chemical composition of the OS bio oil as determined by GCMS (massHunter/library/NIST 14.1) showed the occurrence of octadecadienoic and oleic acids as poly-saturated and saturated fat found in plant glycosides and vegetable fats and oil. The DSC thermal stability analysis showed the possible existence of mixed triglycerides. Finally, the result from the physiochemical characterization compositions, and functional group the OS bio oil indicated its potential as a suitable substitute for petroleum-based oil.

The apiosyltransferase celery UGT94AX1 catalyzes the biosynthesis of the flavone glycoside apiin.

Apiose is a unique branched-chain pentose found in plant glycosides and a key component of the cell wall polysaccharide pectin and other specialized metabolites. More than 1,200 plant-specialized metabolites contain apiose residues, represented by apiin, a distinctive flavone glycoside found in celery (Apium graveolens) and parsley (Petroselinum crispum) in the family Apiaceae. The physiological functions of apiin remain obscure, partly due to our lack of knowledge on apiosyltransferase during apiin biosynthesis. Here, we identified UGT94AX1 as an A. graveolens apiosyltransferase (AgApiT) responsible for catalyzing the last sugar modification step in apiin biosynthesis. AgApiT showed strict substrate specificity for the sugar donor, UDP-apiose, and moderate specificity for acceptor substrates, thereby producing various apiose-containing flavone glycosides in celery. Homology modeling of AgApiT with UDP-apiose, followed by site-directed mutagenesis experiments, identified unique Ile139, Phe140, and Leu356 residues in AgApiT, which are seemingly crucial for the recognition of UDP-apiose in the sugar donor pocket. Sequence comparison and molecular phylogenetic analysis of celery glycosyltransferases suggested that AgApiT is the sole apiosyltransferase-encoding gene in the celery genome. Identification of this plant apiosyltransferase gene will enhance our understanding of the physioecological functions of apiose and apiose-containing compounds.

Isolation and Identification of Phenolic Profiles in Selected Himalayan Wild Berries and Determination of their Antimicrobial Activity

A fundamental source of food and medicine for nearby Himalayan populations is the diversity of plants. The percentage yield of extract, phytochemicals, and minimum inhibitory concentration (MIC) for the extracts Euterpe oleraceae, Vaccinium myrtillu, Phyllanthus embilica, Rubus ellipticus, and Rubus niveus were determined. The clinical and laboratory standards Institute’s micro broth dilution method was used to assess these extracts’ antimicrobial effects on Klebsiella pneumoniae and Pseudomonas aeruginosa, while the purification of the positive extract was done by high-pressure liquid chromatography. Phytochemical analysis was done to determine the secondary plant metabolites, including alkaloids, polyphenols, glycosides, tannins, flavonoids, carbohydrates, steroids, and saponins. All five of the studied plants’ extracts showed antibacterial activity against one or more tested microorganisms. Several phenolic acids (chlorogenic acid, salicylic acid, ellagic acid, ferulic acid, gallic acid, and caffeic acid were detected in all extracts. The R. ellipticus extracts with petroleum ether, chloroform, methanol, and water show a maximum yield between (64–56%) except R. ellipticus extract with hexane (25.82%) showed a low yield. All the extracts have major quantities of carbohydrates, flavonoids, and phenols. These results suggested that produced antimicrobial activity was due to the presence of phytoconstituents in all extracts.

A review on applications of β-glucosidase in food, brewery, pharmaceutical and cosmetic industries

β-glucosidases hydrolyse glycosidic bonds to release non-reducing terminal glucosyl residues from glycosides and oligosaccharides via catalytic mechanisms. It is very well known that the β-glucosidase enzyme is used in biorefineries for cellulose degradation, where β-glucosidases is the rate-limiting enzyme for the final glucose production from cellobiose. The β-glucosidase enzyme is used as a catalyst in other industrial sectors, including pharmaceuticals, breweries, dairy, and food processing. With the aid of β-glucosidase enzymes, cyanogenic glycosides and plant glycosides are transformed into sugar moiety and aglycones. These aglycone compounds are employed as aromatic compounds in the food processing and brewing industries. They are also used as medications and dietary supplements based on their pharmacological qualities. Applications of aglycones and the microbiological sources of β-glucosidase in aglycone production have been discussed in this review.

Isolation and Characterization of Flavonoid Naringenin and Evaluation of Cytotoxic and Biological Efficacy of Water Lilly (Nymphaea mexicana Zucc.).

Despite its limited exploration, Nymphaea mexicana Zucc. can be beneficial if pharmacology, isolation, and biological evaluation are given attention. It is an aquatic species that belongs to the family Nymphaeaceae. The thrust area of the work was the extraction, isolation, and biological evaluation of different extracts of the N. mexicana Zucc. plant. The primary goal of this research was to assess the antimicrobial, antioxidant, and anticancer activities of the extracts and to isolate the target naringenin compound. Comparative FT IR analysis of different extracts of this plant revealed the presence of functional groups of plant secondary metabolites, including polyphenols, flavonoids, terpenoids, esters, amines, glycosides, alkanes, alkaloids, fatty acids, and alcohols. Moderate free radical scavenging potential has been achieved for the various extracts via reducing power and DPPH assays. While cytotoxic activity was evaluated by colorimetric and lactate dehydrogenase cell viability tests on potent cancer cell lines. Lung adenocarcinoma epithelial cells (A-549), and breast cells (MC-7) were treated with MeOH extract. The antimicrobial activity against bacterial strains was evaluated using Gram-positive and -negative cultures, where maximum and minimum inhibition zones were recorded for different strains, including 1.6-25.6 μg/mL for Streptococcus aureus, using the agar well diffusion method. In addition, the anti-inflammatory activity of different extracts of N. mexicana Zucc. was evaluated in a nitrite radical scavenging assay with high concentrations of secondary metabolites, which are important against human pathogens and other diseases.

A Novel 3-O-rhamnoside: 2″-O-xylosyltransferase Responsible for Terminal Modification of Prenylflavonol Glycosides in Epimedium pubescens Maxim.

Prenylated flavonol glycosides in Epimedium plants, as key medicinal components, are known to have great pharmaceutical activities for human health. Among the main prenylated flavonol glycosides, the modification mechanism of different sugar moieties is still not well understood. In the current study, a novel prenylated flavonol rhamnoside xylosyltransferase gene (EpF3R2″XylT) was cloned from E. pubescens, and the enzymatic activity of its decoding proteins was examined in vitro with different prenylated flavonol rhamnoside substrates and different 3-O-monosaccharide moieties. Furthermore, the functional and structural domains of EpF3R2″XylT were analyzed by bioinformatic approaches and 3-D protein structure remodeling. In summary, EpF3R2″XylT was shown to cluster with GGT (glycosyltransferase that glycosylates sugar moieties of glycosides) through phylogenetic analysis. In enzymatic analysis, EpF3R2″XylT was proven to transfer xylose moiety from UDP-xylose to prenylated flavonol rhamnoside at the 2″-OH position of rhamnose. The analysis of enzymatic kinetics showed that EpF3R2″XylT had the highest substrate affinity toward icariin with the lowest Km value of 75.96 ± 11.91 mM. Transient expression of EpF3R2″XylT in tobacco leaf showed functional production of EpF3R2″XylT proteins in planta. EpF3R2″XylT was preferably expressed in the leaves of E. pubescens, which is consistent with the accumulation levels of major prenylflavonol 3-O-triglycoside. The discovery of EpF3R2″XylT will provide an economical and efficient alternative way to produce prenylated flavonol trisaccharides through the biosynthetic approach.

Salicylic acid elicitation improves antioxidant activity of spinach leaves by increasing phenolic content and enzyme levels

Elicitation is used in agriculture as a method of crop protection or a means to enhance nutraceutically or pharmaceutically important metabolites with the help of chemical stimuli known as elicitors. In the present experiment elicitor potential of Salicylic acid has been exploited to induce metabolic changes in spinach plant. Salicylic acid of various concentrations (1–0.01 mg/ml) were applied on spinach leaves through foliar spray and observed for biochemical changes. A multifold increase in the amount of total phenolic and total flavonoid as well as enzyme activity of peroxidase, catalase and superoxide dismutase was observed in a dose dependent manner. An increase in the antioxidant activity was observed and was shown to be correlated with the phenolic compounds. Growth parameters measured in the form of total biomass and total chlorophyll were decreased by high concentrations. Metabolite profile of the control and treated leaves was generated through ESI-MS in the positive and negative ion modes. The results showed an increase in the ion intensity peak of phenolic acids, such as gentisic acid, hydroxycinnamic acid, coumaric acid, ferulic acid and caffeic acid as well as major flavonoids of spinach plant, like kaempferol, quercetin, isorhamnetin and patulitin glycosides. Ferulic and caeffic acids were quantified by HPLC and showed an increase in their amount after elicitor treatment.