Browsing by Subject "Betalains"
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- PublicationOpen AccessAntitumoral drug potential of tryptophan betaxanthin and related plant betalains in the Caenorhabditis elegans tumoral model(MDPI, 2020-07-22) Henarejos Escudero, Paula; Hernández García, Samanta; Guerrero Rubio, M. Alejandra; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular ABetalains are plants pigments identified as potent antioxidant molecules, naturally present in foods like beetroot and prickly pears. Although activities described for betalain-containing formulations include cancer prevention and treatment, the use of extracts instead of purified pigments has avoided the investigation of the real chemopreventive and chemotherapeutic potential of these phytochemicals. Three betalain-rich extracts and six individual pure betalains were used in this work to characterize the activity and to explore possible molecular mechanisms. The animal model Caenorhabditis elegans (tumoral strain JK1466) was used to evaluate the effect of betalains as chemotherapeutics drugs. An objective evaluation method of tumor growth in C. elegans has been developed to assess the possible antitumoral activity of the different treatments. This protocol allowed a fast and reliable screening of possible antitumoral drugs. Among the betalains tested, tryptophan-betaxanthin reduced tumor size by 56.4% and prolonged the animal’s lifespan by 9.3%, indicating high effectiveness and low toxicity. Structure–activity relationships are considered. Assays with mutant strains of C. elegans showed that the mechanism underlying these effects was the modulation of the DAF-16 transcription factor and the insulin signaling pathway. Our results indicate that tryptophan-betaxanthin and related betalains are strong candidates as antitumoral molecules in cancer treatment.
- PublicationOpen AccessAre seven amino acid substitutions sufficient to explain the evolution of high L-DOPA 4,5-dioxygenase activity leading to betalain pigmentation? Revisiting the gain-of-function mutants of Bean et al. (2018)(Wiley, 2023-05-27) Guerrero-Rubio, M. Alejandra; Walker-Hale, Nathanael; Guo, Rui; Sheehan, Hester; Timoneda, Alfonso; Brockington, Samuel F.; Gandía Herrero, Fernando; Bioquímica y Biología Molecular AThis work revisits a publication by Bean et al. (2018) that reports seven amino acid substitu-tions are essential for the evolution of L-DOPA 4,5-dioxygenase (DODA) activity in Caryo-phyllales. In this study, we explore several concerns which led us to replicate the analyses of Bean et al. (2018). Our comparative analyses, with structural modelling, implicate numerous residues addi-tional to those identified by Bean et al. (2018), with many of these additional residues occur-ring around the active site of BvDODAα1. We therefore replicated the analyses of Bean et al.(2018) to re-observe the effect of their original seven residue substitutions in a BvDODAα2 background, that is the BvDODAα2-mut3 variant. Multiple in vivo assays, in both Saccharomyces cerevisiae and Nicotiana benthamiana,did not result in visible DODA activity in BvDODAα2-mut3, with betalain production always10-fold below BvDODAα1. In vitro assays also revealed substantial differences in both cataly-tic activity and pH optima between BvDODAα1, BvDODAα2 and BvDODAα2-mut3 proteins,explaining their differing performance in vivo. In summary, we were unable to replicate the in vivo analyses of Bean et al. (2018), and our quantitative in vivo and in vitro analyses suggest a minimal effect of these seven residues inaltering catalytic activity of BvDODAα2. We conclude that the evolutionary pathway to high DODA activity is substantially more complex than implied by Bean et al. (2018)
- PublicationRestrictedBiosynthesis of a novel polymeric chitosan-betaxanthin and characterization of the first sugar-derived betalains and their effects in the in vivo model Caenorhabditis elegans(Elsevier, 2020-09-28) Guerrero Rubio, María Alejandra; Hernández García, Samanta; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular "A"Betaxanthins are nitrogenous plant pigments belonging to the family of betalains and they are known for their health-promoting effects and fluorescent properties. A novel biotechnological approach in the synthesis of these compounds has allowed the synthesis of high amounts of known betalains and of novel, tailor-made betalains through the condensation of the structural unit - betalamic acid - with amine groups of different compounds. Here we describe the synthesis and characterization of chitosan-betaxanthin, the first fluorescent polymeric betaxanthin which forms nanoparticles and that might combine the fluorescent properties of betalains and the properties of chitosan, a sugar polymer widely used with medical purposes. In addition, glucosamine, the structural unit of chitosan, and its stereoisomer galactosamine were shown to condense in solution with betalamic acid. This produced novel molecules with spectral and in vivo antioxidant and anti-aging properties similar to those of biological betaxanthins, which are the first sugar-derived betaxanthins described.
- PublicationOpen AccessCharacterization of betalains, saponins and antioxidant power in differently colored quinoa (Chenopodium quinoa) varieties(Elsevier, 2017-05-02) Jiménez-Atiénzar, Mercedes; Ibáñez-Tremolada, Martha; Gómez-Pando, Luz Rayda; Cabanes Cos, Juana; Escribano Cebrián, Josefa; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular "A"Quinoa was the traditional grain crop used by the prehispanic civilizations in America. Grains are white, black, yellow, and red-violet and plants are cultivated in vast areas of Peru, Bolivia and Ecuador. The recent description of the betacyanin pigment betanin in red-violet varieties is here further analyzed detecting the presence of amaranthin not previously identified in quinoa grains. Yellow-orange grains are characterized for the first time and up to four different betaxanthins are found to be responsible for this coloration. The native fluorescence of the identified betaxanthins makes the surface of the yellow quinoa grains glow with green fluorescent light. The presence of betalains is correlated with high antioxidant and free radical scavenging activities measured under the FRAP, ABTS and ORAC assays in grain extracts of 29 Peruvian varieties. TEAC equivalence is as high as 44.1 and 47.4 mmol Trolox/kg for the yellow and red-violet varieties analyzed respectively.
- PublicationOpen AccessDevelopment of betalain producing callus lines from colored quinoa varieties (Chenopodium quinoa Willd)(American Chemical Society, 2017-12-14) Henarejos Escudero, Paula; Guadarrama-Flores, Berenice; Guerrero Rubio, María Alejandra; Gómez-Pando, Luz Rayda; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular "A"Betalains are water-soluble plant pigments of hydrophilic nature with promising bioactive potential. Among the scarce edible sources of betalains is the grain crop quinoa (Chenopodium quinoa Willd), with violet, red, and yellow grains being colored by these pigments. In this work, call us cultures have been developed from differently colored plant varieties. Stable callus lines exhibited color and pigment production when maintained on Murashige and Skoog medium supplemented with the plant growth regulators 6-benzylaminopurine (8.88 μM) and 2,4-dichlorophenoxyacetic acid (6.79 μM) with a reduction of the nitrogen source to 5.91 mM. Pigment analysis by HPLC-DAD and ESI-MS/MS fully describes the content of individual pigments in the cell lines and allows the first report on the pigments present in quinoa seedlings. Phyllocactin and vulgaxanthin Iare described as novel pigments in the species and show the potential of C. quinoa culture lines in the production of compounds of nutritional value.
- PublicationOpen AccessFirst Betalain-producing bacteria break the exclusive presence of the pigments in the plant kingdom(American Society for Microbiology, 2019-03-19) Contreras Llano, Luis Eduardo; Guerrero Rubio, María Alejandra; Lozada Ramírez, José Daniel; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular AThe biosynthesis of antioxidant pigments, namely, betalains, was believed to be restricted to Caryophyllales plants. This paper changes this paradigm, and enzyme mining from bacterial hosts promoted the discovery of bacterial cultures producing betalains. The spectrum of possible sources of betalain pigments in nature is broadened by our description of the first betalain-forming bacterium, Gluconacetobacter diazotrophicus. The enzyme-specific step is the extradiol cleavage of the precursor amino acid L-dihydroxyphenylalanine (L-DOPA) to form the structural unit betalamic acid. Molecular and functional work conducted led to the characterization of a novel dioxygenase, a polypeptide of 17.8 kDa with a Km of 1.36 mM, with higher activity and affinity than those of its plant counterparts. Its superior activity allowed the first experimental characterization of the early steps in the biosynthesis of betalains by fully characterizing the presence and time evolution of 2,3- and 4,5- seco-DOPA intermediates. Furthermore, spontaneous chemical reactions are characterized and incorporated into a comprehensive enzymatic-chemical mechanism that yields the final pigments. Several studies have demonstrated the health-promoting effects of betalains due to their high antioxidant capacity and their positive effect on the dose-dependent inhibition of cancer cells and their proliferation. To date, betalains were restricted to plants of the order Caryophyllales and some species of fungi, but the present study reveals the first betalain-producing bacterium, as well as the first steps in the formation of pigments. This finding demonstrates that betalain biosynthesis can be expanded to prokaryotes.
- PublicationOpen AccessFluorescent Bioinspired Protein Labeling with Betalamic Acid. Derivatization and Characterization of Novel Protein-Betaxanthins(2016-10) Francisco, García-Carmona; Mercedes, Jiménez-Atiénzar; Cabanes Cos, Juana; Escribano Cebrián, Josefa; Gandía Herrero, Fernando; Bioquímica y Biología Molecular ABetaxanthins are the water-soluble pigments that bestow yellow coloration to fruits, flowers and roots of plants of the Caryophyllales order and present autofluorescence after excitation with blue light. In this work, the semi-synthesis of betaxanthins derived from macromolecules is achieved for the first time by exploiting the reactivity of amine groups belonging to proteins. The synthesis of protein-betaxanthins is demonstrated by spectrophotometry and HPLC-ESI-TOF-MS mass analysis. The derivatization with betalamic acid was in a ratio 1:1 and yielded protein-betaxanthins yellow in color that exhibited fluorescent properties with a maximum excitation wavelength of 476 nm and a maximum emission wavelength of 551 nm. Moreover, staining can be started from purified betalamic acid or directly from raw red beet root extracts. The novel bioinspired labeling reaction allowed protein detection in conventional fluorescence scanning and imaging systems and opens a new perspective for betalamic acid derived molecules as fluorescent probes with multiple biological applications.
- PublicationEmbargoFormation of carboxylated and decarboxylated betalains in ripening grains of Chenopodium quinoa by a dual dioxygenase(Oxford University Press, Society for Experimental Biology, 2022-04-07) Henarejos Escudero, Paula; Martínez-Rodríguez, Pedro; Gomez-Pando, Luz Rayda; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular "A"Chenopodium quinoa (quinoa) is a pseudo-cereal that forms part of the cultural heritage of Andean countries, and its grains have high nutritional value and potential health benefits. Betalains are nitrogenous water-soluble pigments and bioactive molecules that contribute to these health-promoting properties. Betalains are restricted to plants of the order Caryophyllales, to which quinoa belongs. A new family of betalains has been discovered in the form of unconventional decarboxylated pigments. Here, we show that these pigments accumulate in ripening quinoa grains of fluorescent nature, and are putatively based on a dopamine-cleaving activity. This study describes for the first time the purification and molecular and functional characterization of a 4,5-dopamine extradiol dioxygenase enzyme from plants. It is a monomeric protein with a molecular mass of 34.5 kDa characterized by chromatography, electrophoresis, and time-offlight mass spectrometry. We demonstrate that this key enzyme has a dual function in a square-shaped biosynthetic pathway towards the formation of both carboxylated and decarboxylated pigments. Enzyme kinetic properties are characterized for the production of 6-decarboxy-betalamic acid and 3,4-dihydroxy-l-phenylalanine-derived betalamic acid, the two structural units of plant pigment in nature. The profile of multiple betalains present in quinoa grains has been reproduced in one-pot bioreactors containing the novel enzyme and two competing substrates
- PublicationOpen AccessHealth-promoting potential of betalains in vivo and their relevance as functional ingredients: A review(Elsevier, 2022-02-13) Martínez-Rodriguez, Pedro; Guerrero Rubio, María Alejandra; Henarejos Escudero, Paula; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular "A"Background: Betalains are phytochemicals of relevance to the food industry not only for their natural presence in foods and beverages but also due to their utility as food colorants. However, the use of betalains as functional ingredients has not been extended despite their demonstrated health-promoting properties. The use of betalains as nutraceuticals is an emerging field thanks to the accumulation of scientific evidence on their beneficial effects on health in humans and animal models. Scope and approach: In this review, all the health-promoting effects of betalains published to date are collected and discussed with a focus on their promising use as functional ingredients. All studies on animal models and humans are critically analyzed. Key findings and conclusions: The bioactive properties of betalains have been manifested in rodents, fish, and nematodes. Chemopreventive, antioxidant, and anti-inflammatory activities are some of the effects produced by betalains in vivo. Assays performed in humans remain limited, but their conclusions are highly promising in terms of the health-promoting potential of betalains, supporting the use of these compounds as functional ingredients.
- PublicationOpen AccessIn vitro, in vivo, and in silico evidence for the use of plant pigments betalains as potential nutraceuticals against Alzheimer’s disease(Wiley, 2024-07-24) Martínez-Rodriguez, Pedro; Henarejos Escudero, Paula; Hernández García, Samanta; Gandía Herrero, Fernando; Sánchez Ferrer, Álvaro; Bioquímica y Biología Molecular AAnti-amyloidogenic properties of plant pigments betalains as potential nutraceuticals against Alzheimer’s disease have been screened using 24 pure molecules. Twenty-two betalains reduced amyloid aggregation in vitro, eight of them up to 100%, with IC 50values in the micromolar range. Atomic force and transmission electron microscopy images showed the typical fibrils associated with Alzheimer’s disease and how beta-lains avoid its formation. Neuroprotection after ingestion was supported by in vivo experiments with Caenorhabditis elegans. Indoline-betacyanin was the most effective molecule by significantly improving the chemotactic behavior of the CL2355 strain,a model of Alzheimer’s disease. Furthermore, in-depth molecular docking analyses revealed that the pigments interact with the N-terminal region of the amyloid pep-tide. This work is the most comprehensive study in the field and provides in vitro, in vivo, and in silico evidence for the use of betalains as nutraceuticals of relevance in theprevention of Alzheimer’s disease
- PublicationOpen AccessLight emission in betalains: from fluorescent flowers to biotechnological applications(Cell Press, 2020-02) Guerrero Rubio, María Alejandra; Escribano Cebrián, Josefa; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular AThe discovery of visible fluorescence in the plant pigments betalains revealed the existence of fluorescent patterns in flowers of plants of the order Caryophyllales, where betalains substitute anthocyanins. The serendipitous initial discovery led to a systemized characterization of the role of different substructures on the photophysical phenomenon. Strong fluorescence is general to all members of the family of betaxanthins linked to the structural property that the betalamic acid moiety is connected to an amine group. This property has led to bioinspired tailor-made probes and to the development of novel biotechnological applications in screening techniques or microscopy labeling. Here, we comprehensively review the photophysics, photochemistry, and photobiology of betalain fluorescence and describe all current applications.
- PublicationOpen AccessLineage-specific gene radiations underlie the evolution of novel betalain pigmentation in Caryophyllales(Wiley, 2015-05-13) Brockington, Samuel F.; Yang, Ya; Covshoff, Sarah; Hibberd, Julián M.; Sage, Rowan F.; Wong, Gane K. S.; Moore, Michael J.; Smith, Stephen A.; Gandía Herrero, Fernando; Bioquímica y Biología Molecular "A"Betalain pigments are unique to the Caryophyllales and structurally and biosynthetically dis-tinct from anthocyanins. Two key enzymes within the betalain synthesis pathway have beenidentified: 4,5-dioxygenase (DODA) that catalyzes the formation of betalamic acid andCYP76AD1, a cytochrome P450 gene that catalyzes the formation of cyclo-DOPA. We performed phylogenetic analyses to reveal the evolutionary history of the DODA andCYP76AD1 lineages and in the context of an ancestral reconstruction of pigment states weexplored the evolution of these genes in relation to the complex evolution of pigments inCaryophylalles. Duplications within the CYP76AD1 and DODA lineages arose just before the origin of beta-lain pigmentation in the core Caryophyllales. The duplications gave rise to DODA-a andCYP76AD1-a isoforms that appear specific to betalain synthesis. Both betalain-specific isoformswere then lost or downregulated in the anthocyanic Molluginaceae and Caryophyllaceae. Our findings suggest a single origin of the betalain synthesis pathway, with neofunctionaliza-tion following gene duplications in the CYP76AD1 and DODA lineages. Loss of DODA-a andCYP76AD1-a in anthocyanic taxa suggests that betalain pigmentation has been lost twice inCaryophyllales, and exclusion of betalain pigments from anthocyanic taxa is mediated throughgene loss or downregulation. [Correction added after online publication 13 May 2015: in thelast two paragraphs of the Summary the gene name CYP761A was changed to CYP76AD1.]
- PublicationRestrictedNanoparticles of betalamic acid derivatives with cyclodextrins. Physicochemistry, production characterization and stability(Elsevier, 2020-07-14) Guerrero Rubio, Alejandra; Matencio Durán, Adrián; Gandía Herrero, Fernando; López Nicolás, José Manuel; García Carmona, Francisco; Bioquímica y Biología Molecular AThe use of betalains as bioactive compounds has been widely investigated. However, their use as a functional food requires a more exhaustive study because of their photosensitivity and poor stability. For this reason, this works studies the stabilization of two highly bioactive betalain derivatives, phenylethylamine-betaxanthin (Ph- Bx) and indoline-betacyanin (In-Bc) by nanoencapsulation in natural a modified cyclodextrins (CDs). Methyl- β-CD (Mβ-CD) showed the best results for Ph-Bx while Hydroxypropyl-β-CD (HPβ-CD) was better in the case of In- Bc. In addition, the effect of pH and temperature on encapsulation was studied finding that the encapsulation constant (KF) value increased as the pH decreased in the case of Ph-Bx but not in the case of In-Bc. The KF decreased as the temperatures increased. The thermodynamic parameters, ΔH◦, ΔS◦ and ΔG◦, were also calculated. Molecular docking provided further insight into how the different interactions influenced the encapsulation constant. The results show that the presence of CDs during the biotechnological synthesis of Ph-Bx and In- Bc increased production by 51 and 26%, respectively, and also increased the stability of both derivatives. This study represents a first step towards increasing the production of this type of compound for introduction into functional foods.
- PublicationEmbargoProduction of dihydroxylated betalains and dopamine in cell suspension cultures of Celosia argentea var. plumosa(American Chemical Society, 2015-03-01) Guadarrama-Flores, Berenice; Rodríguez-Monroy, Mario; Cruz-Sosa, Francisco; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular "A"Betalains are plant pigments of hydrophilic nature with demonstrated chemopreventive potential in cancer celllines and animal models. Among the betalains, those containing an aromatic moiety with two free hydroxyl groups possess thestrongest antioxidant and free radical scavenging activities. The betaxanthins dopaxanthin and miraxanthin V and the betacyaninsbetanidin and decarboxy-betanidin are the only natural betalains with catecholic substructures. These four pigments have beenproduced in cell cultures established from hypocotyls of the plant Celosia argentea. Two stable and differentially colored cell lines,yellow and red, were maintained on Murashige and Skoog medium supplemented with the plant growth regulators 6-benzylaminopurine (6.66 μM) and 2,4-dichlorophenoxyacetic acid (6.79 μM). Derived suspension cultures showed increasedproduction of dihydroxylated betalains in the cells and secreted to the medium with a maximum reached after 8 days of culture.In addition, precursor molecules betalamic acid and dopamine, with content up to 42.08 mg/g dry weight, were also obtained.The joint presence of the bioactive betalains together with the production of dopamine and betalamic acid show the ability of cellcultures of C. argentea to become a stable source of valuable phytochemicals
- PublicationRestrictedReversible bleaching of betalains induced by metals and application to the fluorescent determination of anthrax biomarker(Elsevier, 2020-05-01) Guerrero Rubio, M. Alejandra; Martínez Zapata, Joaquín; Henarejos Escudero, Paula; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular ABetalains are antioxidant pigments of plant origin that are found in nature in the order Caryophyllales. They are classified into two groups: betacyanins, which present a violet coloration, and betaxanthins, fluorescent betalains with yellow coloration. In this work, the effect of metals, such as copper and europium, on the spectroscopic properties of betalains is studied. For the first time, the reversibility of spectral changes in fluorescence and absorbance is considered after the addition of chelating substances. These results explain the general bleaching of betalains by metals. The reversibility of the betalain-metal complexes is applied to the fluorescent detection of the presence of spores from Bacillus by the interaction of the complexes with the chelator dipicolinic acid, which is the main and characteristic component of endospores. This fast, economic method allows the detection of spores below the infective dose for anthrax by using a flower pigment present in Mirabilis jalapa.