Investigación

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Investigación contiene los documentos generados por los docentes e investigadores de la Universidad en su labor de investigación. Este depósito tiene como objetivos el almacenamiento, la gestión, la preservación y la difusión de estos documentos, y pretende contribuir al aumento de la visibilidad del trabajo desarrollado por los miembros de la comunidad universitaria.

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Now showing 1 - 17 of 17

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A dopamine-based biosynthetic pathway produces decarboxylated betalains in Chenopodium quinoa
(Oxford University Press, American Society of Plant Biologists, 2021-04-07) Henarejos Escudero, Paula; Contreras-Llano, Luis Eduardo; Lozada-Ramírez, José Daniel; Gomez-Pando, Luz Rayda; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular "A"
Betalains are the nitrogenous pigments that replace anthocyanins in the plant order Caryophyllales. Here, we describe unconventional decarboxylated betalains in quinoa (Chenopodium quinoa) grains. Decarboxylated betalains are derived from a previously unconsidered activity of the 4,5-DOPA-extradiol-dioxygenase enzyme (DODA), which has been identified as the key enzymatic step in the established biosynthetic pathway of betalains. Here, dopamine is fully characterized as an alternative substrate of the DODA enzyme able to yield an intermediate and structural unit of plant pigments: 6-decarboxy-betalamic acid, which is proposed and described. To characterize this activity, quinoa grains of different colors were analyzed in depth by chromatography, time-of-flight mass spectrometry, and reactions were performed in enzymatic assays and bioreactors. The enzymatic-chemical scheme proposed leads to an uncharacterized family of 6-decarboxylated betalains produced by a hitherto unknown enzymatic activity. All intermediate compounds as well as the final products of the dopamine-based biosynthetic pathway of pigments have been unambiguously determined and the reactions have been characterized from the enzymatic and functional perspectives. Results evidence a palette of molecules in quinoa grains of physiological relevance and which explain minor betalains described in plants of the Caryophyllales order. An entire family of betalains is anticipated.
Open Access
Bioactive potential and spectroscopical characterization of a novel family of plant pigments betalains derived from dopamine
(Elsevier, 2022-09-22) Henarejos Escudero, Paula; Martínez Rodríguez, Pedro; Gandía Herrero, Fernando; García Carmona, Francisco; Hernández García, Samanta; Bioquímica y Biología Molecular "A"
With two compounds first discovered in quinoa, an entire novel family of betalain pigments derived from dopamine is obtained and characterized. Betalains are nitrogenous water-soluble pigments and bioactive molecules with health-promoting effects and nutraceutical potential. It was assumed that all betalains contained betalamic acid as a structural unit derived from L-dihydroxyphenylalanine (L-DOPA). However, hitherto ignored compounds derived from dopamine have recently been discovered in nature. Here an entire family of betalains is described as decarboxylated pigments where 6-decarboxy-betalamic acid is the chromophoric and structural unit. This paper shows for the first time the production, purification and characterization of color and fluorescent properties of this novel family of pigments. Antioxidant and anti-aging effects of the just discovered betalains were tested in vivo using the animal model Caenorhabditis elegans. Some of them presented extraordinary properties, being glutamic acid-6-decarboxy-betaxanthin the most fluorescent molecule among both families of betalains. Methionine sulfoxide-6-decarboxy-betaxanthin is described as the most potent betalain in the reduction of oxidative stress in vivo in C. elegans (99.5 % at 25 µM) and dopa-6-decarboxy-betaxanthin increased the lifespan of the animal model up to 7.0 % at 25 µM. These results open new research lines in the search for molecules from plants with health-promoting properties and bioactivities.
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Biosynthesis 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) Gandía Herrero, Fernando; García Carmona, Francisco; Hernández García, Samanta; Guerrero Rubio, María Alejandra; 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.
Open Access
Characterization 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.
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Comparative study of the antioxidant capacity of four stilbenes using ORAC, ABTS+, and FRAP Techniques
(Springer, 2017-03-17) Rodríguez-Bonilla, Pilar; Matencio Durán, Adrián; Gandía Herrero, Fernando; López Nicolás, José Manuel; García Carmona, Francisco; Bioquímica y Biología Molecular "A"
Researchers use several different analytical techniques, such as ORAC, ABTS·+, or FRAP, for measuring the antioxidant capacity of bioactive compounds. However, many authors do not take into account that these three techniques have different objectives. This contribution reports on the use of two types of tests to evaluate the antioxidant activity of four stilbene tests based first on the hydrogen atom transfer reaction (ORAC method) and the second on the single electron transfer reaction (ABTS·+ and FRAP assays). For the ORAC assay, the greatest antioxidant activity was shown by resveratrol, followed by oxyresveratrol, pterostilbene, and pinosylvin, while in the ABTS.+ assay, the highest antioxidant capacity was presented by oxyresveratrol, followed by resveratrol, pinosylvin, and finally pterostilbene. In the FRAP assay, the reducing activity shown by all the stilbenes was below that obtained for trolox. The role of phenolic hydroxyl groups was studied. The technique used should be selected taking into account the objectives and the conditions of the medium.
Open Access
Development of betalain producing callus lines from colored quinoa varieties (Chenopodium quinoa Willd)
(American Chemical Society, 2017-12-14) Henarejos Escudero, Paula; Guadarrama-Flores, Berenice; Gómez-Pando, Luz Rayda; Gandía Herrero, Fernando; García Carmona, Francisco; Guerrero Rubio, María Alejandra; 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.
Open Access
Escherichia coli protein YgiD produces the structural unit of plant pigments betalains: characterization of a prokaryotic enzyme with DOPA-extradiol-dioxygenase activity
(Springer, 2013-05-12) Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular "A"
Betalamic acid is the structural unit of all the natural pigments betalains. These are nitrogen-containing water soluble compounds with high colorant and bioactive properties, characteristic of plants of the order Caryophyllales. The formation of betalamic acid from the precursor amino acid 3,4-dihydroxy-L-phenylalanine (L-DOPA) by the enzyme 4,5- DOPA-extradiol-dioxygenase was supposed to be restricted to plants of this order and two fungal species. Here, the first case of betalamic acid formation by an enzyme other than eukaryotes is reported with a homolog enzyme from Escherichia coli. The protein YgiD has been cloned, expressed, and purified to carry out its molecular and functional characterization. The enzyme was obtained as a monomeric active protein with a molecular mass of 32 kDa characterized by chromatography, electrophoresis, and MALDI-TOF analysis. Enzyme kinetic properties are characterized in the transformation of the relevant substrate L-DOPA. Reaction was analyzed spectrophotometrically and by HPLC-DAD, electrospray ionization mass spectrometry, and time-of-flight mass spectrometry
Open Access
Flavonoids’ effects on Caenorhabditis elegans’ longevity, fat accumulation, stress resistance and gene modulation involve mTOR, SKN-1 and DAF-16
(MDPI, 2021-03-12) Gandía Herrero, Fernando; García Carmona, Francisco; Hernández García, Samanta; Guerrero Rubio, María Alejandra; Bioquímica y Biología Molecular "A"
Flavonoids are potential nutraceutical compounds present in diary food. They are considered health-promoting compounds and promising drugs for different diseases, such as neurological and inflammatory diseases, diabetes and cancer. Therefore, toxicological and mechanistic studies should be done to assert the biological effects and identify the molecular targets of these compounds. In this work we describe the effects of six structurally-related flavonoids—baicalein, chrysin, scutellarein, 6-hydroxyflavone, 6,7-dihydroxyflavone and 7,8-dihydroxyflavone—on Caenorhabditis elegans’lifespan and stress resistance. The results showed that chrysin, 6-hydroxyflavone and baicalein prolonged C. elegans’ lifespan by up to 8.5%, 11.8% and 18.6%, respectively. The lifespan extensions caused by these flavonoids are dependent on different signaling pathways. The results suggested that chrysin’s effects are dependent on the insulin signaling pathway via DAF-16/FOXO. Baicalein and 6-hydroxyflavone’s effects are dependent on the SKN-1/Nfr2 pathway. In addition, microarray analysis showed that baicalein downregulates important age-related genes, such as mTOR and PARP.
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Formation 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
Open Access
Health-promoting potential of betalains in vivo and their relevance as functional ingredients: A review
(Elsevier, 2022-02-13) Martínez-Rodriguez, Pedro; Henarejos Escudero, Paula; Gandía Herrero, Fernando; García Carmona, Francisco; Guerrero Rubio, María Alejandra; 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.
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Inactivation of lipoxygenase and cyclooxygenase by natural betalains and semi-synthetic analogues
(Elsevier, 2014-01-13) Vidal, Pedro J.; Gandía Herrero, Fernando; López Nicolás, José Manuel; García Carmona, Francisco; Bioquímica y Biología Molecular "A"
Betalains are natural pigments characteristic of plants of the order Caryophyllales. In this work, the role of betalains in the anti-inflammatory activity described for plant extracts is analysed in terms of the inactivation of the enzymes involved in the biochemical response (lipoxygenase and cyclooxygenase). Pure natural betalains and semi-synthetic analogues are demonstrated to promote a significant reduction of the enzymes activity. Reactions were followed spectrophotometrically and by HPLC-DAD. Phenethylamine-betaxanthin was the most potent in the inactivation of cyclooxygenase, with a reduction of 32% of the control activity at 125 lM, while the natural pigment betanidin and a betalain analogue derived from indoline resulted as the most potent inactivators of lipoxygenase, with IC50 values of 41.4 and 40.1 lM, respectively. Molecular docking studies revealed that betalains interact with the lipoxygenase amino acids involved in substrate binding and with Tyr-385 and Ser-530 close to the yclooxygenase active site, interfering in enzyme catalysis.
Open Access
Lineage-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.]
Open Access
Modifying the chromophoric ring of betalamic acid in plant pigments. Synthesis and characterization of methylated betalains
(Elsevier, 2025-05-23) Martínez-Rodríguez, Pedro; Contreras-Llano, Luis Eduardo; Pagán-López, Diego José; Lozada-Ramírez, José Daniel; Gandía Herrero, Fernando; Hernández García, Samanta; Guerrero Rubio, María Alejandra; Bioquímica y Biología Molecular "A"
The recent discovery of decarboxylated betalains, derived from dopamine, in nature has expanded the number of chemical modifications possible in these plant pigments. Alternative starting materials in biochemical rocesses can yield novel molecules with modifications in the betalamic ring. For the first time, a functional group has been incorporated into the betalamic acid structure of betalains, thus creating a novel family of methylated pigments. A synthetic pathway starting from α-methyl-DOPA yields 6-methyl-betalamic acid. The process is enzymatic and kinetic characterization reveals that the formation of 6-methyl-betalamic acid is favored over the formation of 6- methyl-dopaxanthin, as a further condensation reaction can be hindered. Starting from the methylated betalamic acid, a novel family of methylated betalains has been biotechnologically produced comprising ten yellow and violet molecules. The novel compounds were characterized by using HPLC-ESI-Q-TOF-MS, spectrophotometry, and spectrofluorometry. Potential antioxidant capacity was measured using the ORAC assay, demonstrating the molecules’ effectiveness against peroxyl radicals, with indoline-6-methyl-betacyanin identified as the most antioxidant. This study shows the venue for the chemical modification of the betalamic ring in pigments and thus represents the beginning of the era of betalain core chemistry. This opens new fields for future research to explore novel structures and their potential bioactive effects.
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One-Step synthesis of betalains using a novel betalamic acid derivatized support
(American Chemical Society, 2014-04-01) Jiménez-Atiénzar, Mercedes; Cabanes Cos, Juana; Escribano Cebrián, Josefa; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular "A"
Betalains are plant pigments with high antioxidant and cancer hemopreventive properties used by the food industry as safe colorants. Betalains are restricted to species of the order Caryophyllales, and difficulty in obtaining individual molecules has limited their structural identification and application. This study was designed to develop a betalamic acid derivatized support generated from a primary amine polymer. The novel material presents color properties of a pseudobetaxanthin, and it is stable for at least 6 months. The bond formed can be displaced at mild conditions by the addition of amines in aqueous solutions over a broad pH range and at 25 °C. This releases the betalamic acid while forming the corresponding pigment. This one-step procedure significantly simplifies the process of obtaining semisynthetic betalains, and it is optimized here for the formation of betaxanthins and betacyanins derived from tyramine, dopamine, pyrrolidine, and indoline. The new method makes access to single betalains available to the entire scientific community and could stimulate research and applications in the field.
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Polyphenols from traditional Chinese medicine and Mediterranean diet are effective against Aβ toxicity in vitro and in vivo in Caenorhabditis elegans
(Royal Society of Chemistry, 2021-12-15) Gea-González, Adriana; Henarejos Escudero, Paula; Martínez-Rodríguez, Pedro; Gandía Herrero, Fernando; García Carmona, Francisco; Hernández García, Samanta; Bioquímica y Biología Molecular "A"
The potential of naturally occurring polyphenols as nutraceuticals to prevent and/or treat Alzheimer’s disease is studied. Five structurally related flavones and four tyrosols were tested in vitro in human amyloid-β peptide aggregation assays. The most promising compounds were two flavones, scutellarein and baicalein, and two tyrosols hydroxytyrosol and hydroxytyrosol acetate. These compounds caused a dose-dependent reduction of Aβ-peptide aggregation up to 90% for the flavones and 100% for the tyrosols, at concentrations of 83.3 µM and 33.3 mM, respectively. The IC50 value obtained for scutellarein was 22.5 µM, and was slightly higher for baicalein, 25.9 µM, while for hydroxytyrosol and hydroxytyrosol acetate they were 0.57 mM and 0.62 mM. Given these results, the compounds were selected to conduct in vivo assays with the Caenorhabditis elegans animal model of Alzheimer’s disease. The amyloid antiaggregation ability of these polyphenols was demonstrated in in vivo aggregation assays in which 1 mM hydroxytyrosol reduced the amyloid plaques in the mutant strain CL2331 by 43%. The neuroprotective effect was evaluated in chemotaxis experiments carried out with transgenic strain CL2355 that expresses the human amyloid-β peptide in the neurons. The chemotaxis index was improved by 240% when the neuron-impaired animals were treated with 1 mM hydroxytyrosol. The results indicate that the four molecules would be viable candidates to develop nutraceuticals that interfere in amyloid-β peptide aggregation and, consequently, prevent and/or treat Alzheimer’s disease.
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Production 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
Open Access
Scaled-up biotechnological production of individualbetalains in a microbial system
(Wiley, 2019-07-04) López-Llorca, Rosalía; Henarejos Escudero, Paula; Gandía Herrero, Fernando; García Carmona, Francisco; Guerrero Rubio, María Alejandra; Bioquímica y Biología Molecular "A"
The recent interest in plant pigment betalains asbioactive compounds and chemopreventive agents has led to the search for a reliable and scalable process to obtain them. The cloning of the novel andefﬁcient enzyme 4,5-DOPA-extradiol dioxygenasefrom Gluconacetobacter diazotrophicus in an expres-sion vector, and the subsequent heterologous expression in Escherichia coli cultures has led to the start-up of a biotechnological production system of individual pigments. The aim of this study was to search for the optimal conditions for the production of betalamic acid in microbial factories and the scaled-up obtention of the derived pigments. Four different betaxanthins and two betacyanins were obtained after the addition of non-transformable amines and amino acids and their condensation with thebetalamic acid produced by the dioxygenase. The scaled-up obtention and puriﬁcation of betalains improved the yields of the previous methodologies reaching quantities by up to 150 mg of pure compounds.

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