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7 Figuras.-- 3 Tablas, Olive oil production using the two-phase extraction system generates large amounts of biomass named alperujo, which has high percentage of water, slightly acidic pH, and high concentration of salts, organic matter, and phenolic compounds. Due to the presence of these components, alperujo is a significant source of environmental pollution, hindering the disposal of this waste. Nevertheless, some phenolic compounds such as tyrosol and hydroxytyrosol, have high nutraceutical and antioxidant power and could have potential applications in the food, pharmaceutical and cosmetic industries. This work aimed to evaluate the ability of different alperujo-derived media, with different levels of phenolic compounds, to sustain the growth and carotenoid production by yeasts from Rhodotorula genera (R. mucilaginosa LPSC 638, R. glutinis Y54, and R. graminis Y52), as well as to prospect the biotechnological production of carotenoids in these media. The yeasts grew and produced carotenoids in all media, although growth was better in dephenolized media. The highest amount of carotenoids (43.0 ± 3.1 mg/g dry yeast) was obtained with R. glutinis Y54 grown in a thermally treated and dephenolized medium. Our study represents a novel approach to valorize alperujo with a dual benefit, the recovery of valuable compounds, i. e. phenolic compounds, and the bioproduction of new ones, i.e. carotenoids., This work was partially supported by “Agencia Nacional de Promoción Científica y Tecnológica” (ANPCyT, PICT 2018-4065), Argentina. VB stay at the Instituto de la Grasa was financed by an External Fellowship from “Consejo Nacional de Investigaciones Científicas y Técnicas” (CONICET), Argentina.

34 Páginas.-- 6 Figuras.-- 2 Tablas, Carotenoids are diet-based micronutrients important in health and coloration signaling. Related species with similar diets can differ in the kinds and levels of circulating carotenoids, which suggests specific physiological mechanisms to efficiently utilize these micronutrients, regardless of their availability. We explored whether diet and parental provisioning of unusual sources of carotenoids (fresh vegetal matter and vertebrate feces) can explain the occurrence and concentrations of carotenoids in the cinereous vulture Aegypius monachus, griffon vulture Gyps fulvus, and Egyptian vulture Neophron percnopterus nestlings, even when these pigments appear to not be deposited in their integumentary system. A greater diversity of wild prey in diet could be behind the profile of higher concentrations of carotenoids in the Egyptian vulture, the species with carotenoid-dependent coloration during adulthood, while differences in diet composition between cinereous and griffon vultures do not translate to different carotenoid profiles. The carotenoid profile appears to not be related to the ingestion of unusual matter rich in these compounds, although the infrequent occurrence of lycopene and an unidentified γ-carotene-like compounds suggest that these vultures may be exploiting vegetal matter that left no identifiable unconsumed remains in the nest of Egyptian vultures. The consumption of green plant material by griffon vultures does not result in especially high levels of carotenoids when compared to the carotenoids found in cinereous vultures, which do not consume green plant material. Ungulate feces were not provisioned to Egyptian vulture nestlings, despite the fact they contain carotenoids that adults need for appropriate coloration. Overall, this study indicates that diet differences alone appear insufficient to explain contrasting inter-specific carotenoid profiles, especially since all types of food consumed are considered to be poor in carotenoids, except vegetable matter. We suggest that nestling Egyptian vultures are comparatively efficient in up taking carotenoids present in low concentrations in food when these compounds are not deposited in their integument, which suggests allocation to other functions., We thank J.L. González del Barrio, Ó. Frías, B. Arroyo, F. Martínez, J.M. García, A. González, J.P. Calle, J. García,J. Oltra and M. Carrete for their help with fieldwork, and R. Sebastián for assistance in pigment extractions. J.de la Cruz, A. González and J. García kindly provided pictures. Research was funded through projects CGL2007-61395, CGL2010-15726, CGL2013-42451-P, and PID2019-109685GB-I00 of the Spanish Ministry of Science and Innovation

Carotenoid esterification is a new target for cereal biofortification since esterification increases both accumulation and stability of carotenoids. A xanthophyll acyl transferase is responsible for carotenoid esterification in the endosperm of wheat and related cereals. In this chapter we describe the procedures for transferring the carotenoid esterification attribute into wheat using the wild barley Hordeum chilense as donor of the esterification trait, the outline of the breeding program and the protocols for marker assisted selection and the analysis of carotenoids in grain. Biofortified cereals with increased lutein ester content will help to reduce the risk of developing age-related macular degeneration in human populations with limited access to other dietary sources.

3 Figuras.-- 2 Tablas, Carotenoid esterification is a new target for cereal biofortification since esterification increases both accumulation and stability of carotenoids. A xanthophyll acyl transferase is responsible for carotenoid esterification in the endosperm of wheat and related cereals. In this chapter we describe the procedures for transferring the carotenoid esterification attribute into wheat using the wild barley Hordeum chilense as donor of the esterification trait, the outline of the breeding program and the protocols for marker assisted selection and the analysis of carotenoids in grain. Biofortified cereals with increased lutein ester content will help to reduce the risk of developing age-related macular degeneration in human populations with limited access to other dietary sources., This research was funded by project AGL2017-85368-P and by MCIN/AEI/10.13039/501100011033/FEDER “Una Manera de hacer Europa.” M.D.R.-R. was supported by PRE2018-084037 funded by MCIN/AEI/10.13039/501100011033 and FSE “EL FSE invierte en tu futuro.”

Carotenoids are isoprenoids widely distributed in foods that have been always part of the diet of humans. Unlike the other so-called food bioactives, some carotenoids can be converted into retinoids exhibiting vitamin A activity, which is essential for humans. Furthermore, they are much more versatile as they are relevant in foods not only as sources of vitamin A, but also as natural pigments, antioxidants, and health-promoting compounds. Lately, they are also attracting interest in the context of nutricosmetics, as they have been shown to provide cosmetic benefits when ingested in appropriate amounts. In this work, resulting from the collaborative work of participants of the COST Action European network to advance carotenoid research and applications in agro-food and health (EUROCAROTEN, www.eurocaroten.eu, https://www.cost.eu/actions/CA15136/#tabs|Name:overview) research on carotenoids in foods and feeds is thoroughly reviewed covering aspects such as analysis, carotenoid food sources, carotenoid databases, effect of processing and storage conditions, new trends in carotenoid extraction, daily intakes, use as human, and feed additives are addressed. Furthermore, classical and recent patents regarding the obtaining and formulation of carotenoids for several purposes are pinpointed and briefly discussed. Lastly, emerging research lines as well as research needs are highlighted., This article is based upon work from COST Action (European network to advance carotenoid research and applications in agro-food and health, EUROCAROTEN, CA15136, www.eurocaroten.eu, https://www.cost.eu/actions/CA15136/#tabs|Name:overview) supported by COST (European Cooperation in Science and Technology, http://www.cost.eu/).

The Salt-Overly-Sensitive (SOS) pathway controls the net uptake of sodium by roots and the xylematic transfer to shoots in vascular plants. SOS3/CBL4 is a core component of the SOS pathway that senses calcium signaling of salinity stress to activate and recruit the protein kinase SOS2/CIPK24 to the plasma membrane to trigger sodium efflux by the Na/H exchanger SOS1/NHX7. However, despite the well-established function of SOS3 at the plasma membrane, SOS3 displays a nucleo-cytoplasmic distribution whose physiological meaning is not understood. Here, we show that the N-terminal part of SOS3 encodes structural information for dual acylation with myristic and palmitic fatty acids, each of which commands a different location and function of SOS3. N-myristoylation at glycine-2 is essential for plasma membrane association and recruiting SOS2 to activate SOS1, whereas S-acylation at cysteine-3 redirects SOS3 toward the nucleus. Moreover, a poly-lysine track in positions 7–11 that is unique to SOS3 among other Arabidopsis CBLs appears to be essential for the correct positioning of the SOS2-SOS3 complex at the plasma membrane for the activation of SOS1. The nuclear-localized SOS3 protein had limited bearing on the salt tolerance of Arabidopsis. These results are evidence of a novel S-acylation dependent nuclear trafficking mechanism that contrasts with alternative subcellular targeting of other CBLs by S-acylation

10 Páginas.-- 4 Figuras.-- 1 Tabla, Carotenoids are essential in the human diet for their important functions in health. Besides, they are responsible for the yellow pigments desirable for industrial quality in durum wheat. The remarkable carotenoid content of durum wheat endosperm is mostly due to lutein. However, lutein esters have not been previously detected in durum wheat as in other cereals such as common wheat, tritordeum or Hordeum chilense. Esterification increases carotenoid stability and allows greater retention and accumulation through the food chain. Therefore, carotenoid esterification is revealed as a new key target in breeding. We characterized the carotenoid profile of 156 accessions of the Spanish durum wheat collection, searching for landraces with esterification ability. Interestingly, four accessions produced lutein monoesters and diesters. Also, traces of lutein monoesters were detected in eleven accessions. The identification of the first durum wheat accessions with esterification ability reported herein is a remarkable advance for carotenoid biofortification. Furthermore, variation for the relative content of zeaxanthin, α-carotene and β-carotene was also observed. This diversity for the β,ε and β,β branches of the carotenogenic pathway also represents a new opportunity for breeding for specific carotenoids in biofortification programs., This research was funded by Ministerio de Ciencia, Innovación y Universidades, grant number AGL2017-85368-P and by Instituto de Investigación y Formación Agraria y Pesquera (IFAPA), Junta de Andalucía, grant number PP.AVA-AVA2019.020. Both grants are cofounded by FEDER. D.H.-M. is member of CaRed Network funded by Ministerio de Economía y Competitividad (BIO2017-90877-REDT) and participates in EUROCAROTEN COST Action (CA15136). S.G.A. is member of FiRCMe Network, funded by Ministerio de Economía, Industria y Competitividad (AGL2016-81855-REDT). M.D.R.-R. was recipient of a fellowship from Ministerio de Ciencia, Innovación y Universidades (PRE2018-084037).

6 Páginas.-- 6 Tablas, This study was aimed at making progress on the valorisation of table olive wastewater that currently represent a big environmental problem for factories. Concentrates from vacuum evaporation of the wastewater generated during processing of black ripe olives treated with KOH were tested as fertilisers of tomato plants in open field assays for three consecutive crops. Fertilisation was performed by drip irrigation every 15 days; the first treatment being 15 days after transplanting, and a total of five fertilisation treatments were carried out. A phytotoxic effect was not observed on plants or fruit in any case but higher yield (fruit/plant and g/plant) was obtained in comparison to irrigation with only tap water. Moreover, the combined use of the olive concentrate with inorganic nitrogen supply (NH4NO3), in order to comply with the nutrient needs of plants, was also tested. The results demonstrated that the olive concentrates could be a good substitute for inorganic potassium (as KNO3) during the cultivation of tomato plants without any negative effect on the tomato quality (pH, °Brix, sugars, organic acids) or content of bioactive substances (phenolic compounds and carotenoids)., This work was supported by the Spanish Government (Project AGL2016-76820-R, AEI/FEDER, UE). We thank Alejandra Expósito for her technical assistance.

Carotenoids are isoprenoids widely distributed in foods that have been always part of the diet of humans. Unlike the other so-called food bioactives, some carotenoids can be converted into retinoids exhibiting vitamin A activity, which is essential for humans. Furthermore, they are much more versatile as they are relevant in foods not only as sources of vitamin A, but also as natural pigments, antioxidants, and health-promoting compounds. Lately, they are also attracting interest in the context of nutricosmetics, as they have been shown to provide cosmetic benefits when ingested in appropriate amounts. In this work, resulting from the collaborative work of participants of the COST Action European network to advance carotenoid research and applications in agro-food and health (EUROCAROTEN, www.eurocaroten.eu, https://www.cost.eu/actions/CA15136/#tabs|Name:overview) research on carotenoids in foods and feeds is thoroughly reviewed covering aspects such as analysis, carotenoid food sources, carotenoid databases, effect of processing and storage conditions, new trends in carotenoid extraction, daily intakes, use as human, and feed additives are addressed. Furthermore, classical and recent patents regarding the obtaining and formulation of carotenoids for several purposes are pinpointed and briefly discussed. Lastly, emerging research lines as well as research needs are highlighted.

Carotenoids are essential in human diet, so that the development of programs toward carotenoid enhancement has been promoted in several crops. The cereal tritordeum, the amphiploid derived from the cross between Hordeum chilense Roem. et Schulz. and durum wheat has a remarkable carotenoid content in the endosperm. Besides, a high proportion of these carotenoids are esterified with fatty acids. The identification of the gene(s) responsible for xanthophyll esterification would be useful for breeding as esterified carotenoids show an increased ability to accumulate within plant cells and have a higher stability during post-harvest storage. In this work, we analyzed five genes identified as candidates for coding the xanthophyll acyltransferase (XAT) enzyme responsible for lutein esterification in H. chilense genome. All these genes were expressed during grain development in tritordeum, but only HORCH7HG021460 was highly upregulated. Sequence analysis of HORCH7HG021460 revealed a G-to-T transversion, causing a Glycine to Cysteine substitution in the protein of H290 (the only accession not producing quantifiable amounts of lutein esters, hereinafter referred as zero-ester) of H. chilense compared to the esterifying genotypes. An allele-specific marker was designed for the SNP detection in the H. chilense diversity panel. From the 93 accessions, only H290 showed the T allele and the zero-ester phenotype. Furthermore, HORCH7HG021460 is the orthologue of XAT-7D, which encodes a XAT enzyme responsible for carotenoid esterification in wheat. Thus, HORCH7HG021460 (XAT-7Hch) is a strong candidate for lutein esterification in H. chilense and tritordeum, suggesting a common mechanism of carotenoid esterification in Triticeae species. The transference of XAT-7Hch to wheat may be useful for the enhancement of lutein esters in biofortification programs., This research was funded by grant AGL2017-85368-P from Ministerio de Ciencia, Innovación y Universidades, and co-funded by FEDER. MR-R was recipient of a fellowship from Ministerio de Ciencia, Innovación y Universidades (PRE2018-084037). DH-M is member of CaRed Network, funded by Ministerio de Economía y Competitividad (BIO2017-90877-REDT) and participated in EUROCAROTEN COSTAction (CA15136). SA is member of FiRCMe Network, funded by Ministerio de Economía, Industria y Competitividad (AGL2016-81855-REDT).

Neurosporaxanthin (NX) is a carboxylic carotenoid produced by some filamentous fungi, including species of the genera Neurospora and Fusarium. NX biosynthetic genes and their regulation have been thoroughly investigated in Fusarium fujikuroi, an industrial fungus used for gibberellin production. In this species, carotenoid-overproducing mutants, affected in the regulatory gene carS, exhibit an upregulated expression of the NX pathway. Based on former data on a stimulatory effect of nitrogen starvation on carotenoid biosynthesis, we developed culture conditions with carS mutants allowing the production of deep-pigmented mycelia. With this method, we obtained samples with ca. 8 mg NX/g dry mass, in turn the highest concentration for this carotenoid described so far. NX-rich extracts obtained from these samples were used in parallel with carS-complemented NX-poor extracts obtained under the same conditions, to check the antioxidant properties of this carotenoid in in vitro assays. NX-rich extracts exhibited higher antioxidant capacity than NX-poor extracts, either when considering their quenching activity against [O2(1Δg)] in organic solvent (singlet oxygen absorption capacity (SOAC) assays) or their scavenging activity against different free radicals in aqueous solution and in liposomes. These results make NX a promising carotenoid as a possible feed or food additive, and encourage further studies on its chemical properties.

24 Páginas.-- 1 Figura.-- 7 Tablas, Statistical experimental designs were used to formulate a culture medium for zeaxanthin production by an Antarctic Flavobacterium sp. P8 strain. Eleven nutritional factors were assayed in shaken flasks. The effect of temperature on zeaxanthin and carotenoid production was also studied. Peptone, yeast extract, and sodium chloride were the nutrients that caused the principal impact on the biomass growth. These components were further studied to enhance zeaxanthin and total carotenoid concentrations. Although a high production rate of zeaxanthin and carotenoids was achieved, the aerobic characteristics of the bacterial strain and the oxygen requirements for zeaxanthin biosynthesis incorporate a factor that requires additional consideration. Scaling up the process to a 5 L-bioreactor that increased dissolved oxygen availability resulted in a 4.5-fold increase in the total carotenoid content and an almost 9-fold increase in zeaxanthin, which represented 98% of the total carotenoids produced. The results reveal that Flavobacterium sp. P8 is a promising strain for zeaxanthin production., The authors would like to thank Comisión Académica de Posgrado (CAP) for the postgraduate scholarship given to Eugenia Vila and the Instituto Antártico Uruguayo and Comisión Sectorial de Investigación Científica (CSIC) for their financial support (CSIC I+D 2014 219, CSIC INI 230).

36 Páginas.-- 3 Figuras.-- 3 Tablas, Carotenoids are pigmented compounds acquired through diet that have important functions as antioxidants and immune modulators. We studied the association between immunity and circulating carotenoids in Andean condors (Vultur gryphus ). We evaluated the relationship between α‐, β‐, and γ‐globulin blood concentrations and different circulating carotenoids in two groups of Andean condors that differ in their mean health status, rehabilitating (suffering different pathologies), and wild individuals (trapped when displaying their physiological behavior). In rehabilitating individuals, α‐, β‐, and γ‐globulin concentrations were higher than in wild individuals. This shows that rehabilitating individuals were developing an immune response associated with the pathologies that they were suffering at the time of sampling. In addition, circulating carotenoids were lower in rehabilitating than in wild individuals. We found negative correlations between α‐, β‐, and γ‐globulins and different circulating carotenoids in rehabilitating individuals, but not in wild condors. Xanthophylls were strongly related to α‐, β‐, and γ‐globulin blood concentrations in rehabilitating, but not in wild condors. Our results suggest that there is a potential relationship between circulating carotenoids and immunity in the Andean condor. Given that this species may display a carotenoid‐based pigmentation, our results could suggest that a trade‐off between the immune system and external coloration could operate in this species, which may have implications in their access to food resources and mate selection and, thus, in their conservation., We thank the Buenos Aires Zoo staff and Fundación Bioandina Argentina for taking care of condor individuals. We thank P. Alarcón for their comments and suggestions that improved previous versions of this manuscript. We also thank Toni Sanchez‐Zapata, O. Mastrantuoni, M. de la Riva, and the GrInBiC (https://grinbic.com/) team for their help in the field. This study was supported by Agencia Nacional de Promoción Científica y Tecnológica, PICT (BID) 0725‐2014, Fundación BBVA (BIOC‐08‐126), and Universidad Nacional del Comahue project 04/B227. Dámaso Hornero‐Méndez is a member of the Spanish Carotenoid Network (CaRed), funded by MINECO (Spanish Ministry of Economy, Industry, and Competitiveness) projects BIO2015‐71703‐REDT and BIO2017‐90877‐REDT, and the European Carotenoid Network (Euro Caroten; https://www.eurocaroten.eu/), funded by the European Commission COST action CA15136.

Hornero-Méndez D. (2020) A Routine Method for the Extraction and HPLC-DAD Profiling of Major Plant and Food Carotenoids. In: Rodríguez-Concepción M., Welsch R. (eds) Plant and Food Carotenoids. Methods in Molecular Biology, vol 2083. Humana, New York, NY, Carotenoids are ubiquitously present in Nature, and especially in plants and derived foods. The carotenoid profiling is necessary to understand relevant aspects in relation to their biochemistry and genetics, as well as their important roles on human health and animal ecophysiology. Here we propose a simple methodology for the routine extraction and efficient HPLC separation of the most common plant and food carotenoids from plant and animal origins., The author thanks to the Spanish Government (projects AGL2017-85368P and CaRed Network BIO2015-71703-REDT and BIO2017-90877-REDT) for financial support.

BACKGROUND: Alcoholic fermentation of fruits has generated novel products with high concentrations of bioactive compounds and moderate alcohol content. The aim of this study was to evaluate the potential effect on cardiovascular risk factors of the regular consumption by healthy humans of a beverage obtained by alcoholic fermentation and pasteurization of orange juice. RESULTS: Thirty healthy volunteers were enrolled in a randomized controlled study. The experimental group (n = 15) drank 500 mL orange beverage (OB) per day for 2 weeks (intervention phase), followed by a 3-week washout phase. Blood samples were collected at baseline (E-T0) and at the end of the intervention (E-T1) and washout (E-T2) phases. Controls (n = 15) did not consume OB during a 2-week period. OB intake significantly increased oxygen radical absorbance capacity (43.9%) and reduced uric acid (−8.9%), catalase (CAT) (−23.2%), thiobarbituric acid reactive substances (TBARS) (−30.2%) and C-reactive protein (−2.1%) (E-T1 vs. E-T0). These effects may represent longer-term benefits, given the decreased uric acid (−8.9%), CAT (−34.6%), TBARS (−48.4%) and oxidized low-density lipoprotein (−23.9%) values recorded after the washout phase (E-T2 vs. E-T0). CONCLUSION: The regular consumption of OB improved antioxidant status and decreased inflammation state, lipid peroxidation and uric acid levels. Thus OB may protect the cardiovascular system in healthy humans and be considered a novel functional beverage. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry

Carotenoids are ubiquitously present in Nature, and especially in plants and derived foods. The carotenoid profiling is necessary to understand relevant aspects in relation to their biochemistry and genetics, as well as their important roles on human health and animal ecophysiology. Here we propose a simple methodology for the routine extraction and efficient HPLC separation of the most common plant and food carotenoids from plant and animal origins.

3 Figuras, Tritordeums show a significant proportion of lutein esters which increases carotenoid stability and retention throughout the food chain. Esterification is a common means of carotenoid sequestration. A putative association between lutein esters formation acting as a metabolic sink during early stages of grain development and the high carotenoid content of tritordeums is analyzed in this work. Compared to wheat, tritordeums accumulated significantly higher lutein contents from 20 days post anthesis (dpa) but lutein esters were not detected until 36 dpa. Thus esterification is not acting as a metabolific sink before 36 dpa. The presence of lutein esters at late stages of grain development may have a complementary role in carotenoid accumulation by reducing and/or counteracting their catabolism. The differences for lutein esterification among tritordeums suggest the existence of diversity for xanthophyll acyl transferases that could be exploited to increase lutein retention in this cereal and through the food chain., This research was funded by Grants AGL2014-53195-R from Ministerio de Economía y Competitividad and AGL2017-85368-P from Ministerio de Ciencia, Innovación y Universidades. Both projects are cofounded by FEDER. DHM is member of CaRed Network funded by Ministerio de Economía y Competitividad (BIO2015-71703-REDT and BIO2017-90877-REDT) and participates in EUROCAROTEN COST Action (CA15136). SGA is member of FiRCMe Network, funded by Ministerio de Economía, Industria y Competitividad (AGL2016-81855-REDT). M.G.M. was recipient of a fellowship from Ministerio de Economía y Competitividad (BES-2012-055961).

The high carotenoid content and distinctive carotenoid profile of tritordeum are conferred by its wild progenitor, Hordeum chilense. Genetic studies on this wild barley could exploit the knowledge gained in Hordeum vulgare L. if the synteny between H. vulgare and H. chilense is established. DArTSeq markers were aligned to barley genome and used to inspect H. chilense-barley synteny. All chromosome pairs showed a good degree of collinearity with the exception of 7Hv–7Hch, where a reciprocal translocation in 7Hch was identified. Carotenoid analyses revealed a high diversity for total carotenoids, free and esterified lutein in a collection of H. chilense. Population structure analyses revealed the existence of two subgroups contrasting for total carotenoids, free lutein and esterified lutein in seeds. Lutein esters were produced with palmitic and linoleic acids as happens in tritordeum. However, tritordeum prefer palmitic acid for the synthesis of lutein esters but this preference is not maintained in H. chilense. This indicates the existence of diversity in the enzymes involved in the esterification which could be useful in tritordeum breeding. Furthermore, several accessions produced lutein monoesters but they lacked diesters which suggests that esterification is controlled by more than one enzyme in H. chilense. A total of 91 marker-trait associations were identified for carotenoid content and profile. These associations constitute a good starting point for future genetic analyses for the identification of candidate genes from H. vulgare genome., Research funded by Grant AGL2014-53195R, from Ministerio de Economía y Competitividad, Spain (MINECO) including FEDER funding. M.G.M. was recipient of FPI (BES-2012-055961). D.H.-M. is a member of CaRed Network, funded by MINECO (BIO2015-71703-REDT). S.G.A. and CMA are members of FiRCMe Network, funded by MINECO (AGL2016-81855-REDT).

European populations of black-necked grebes (Podiceps nigricollis) congregate every year to moult at the salt ponds of the Odiel Marshes (SW Spain). However, the Odiel Marshes are part of one of the most metal-polluted coastal estuaries in the world, which may pose risks to wildlife. We assessed the exposure of grebes to metal pollution during the critical moulting period in the Odiel Marshes and its potential to cause adverse health effects. Levels of metals in red blood pellet (as a biomarker of exposure), plasma carotenoids, eye redness, and body condition (as biomarkers of effects) were studied. Metal content was also analyzed in the brine shrimp Artemia parthenogenetica, the most important food for grebes in this hypersaline ecosystem during the moulting period. Results showed that, in comparison to toxicity thresholds, grebes had relatively high blood levels of arsenic (As), mercury (Hg) and zinc (Zn). The high loads found in Artemia and the way blood levels vary during the moulting period indicate that shrimp consumption may be the main route of metal exposure for grebes. Plasma carotenoids and body condition showed a positive association with exposure to As, while the relationship of lutein-like carotenoids with Hg accumulation was negative at the beginning of the moulting period to become positive afterwards. Moreover, eye redness was negatively affected by As accumulation. Factors including food resource availability, seasonal fluctuations in physiological status, and interannual variations in the degree of environmental contamination should be considered in monitoring efforts when using moult migrant waterbirds as sentinel species., J. Rodríguez-Estival was supported by a Research and Innovation Strategy for Smart Specialisation (RIS3) postdoctoral contract (2017/11747) from the Junta de Comunidades de Castilla-La Mancha. The study was financially supported by Junta de Andalucía (Consejería de Innovación, Ciencia y Empresa, Project P07-CVI-02700).

Carotenoids are widespread isoprenoids that intervene in actions ranging from the collection of light and photoprotection to the regulation of gene expression and communication within or between species, to mention only some examples. They are, therefore, much more than natural pigments, as they are versatile compounds that are eliciting increasing interest in different disciplines, such as plant science, agriculture, food science and technology, nutrition and health, among others. Although carotenoids in humans are found almost exclusively free, in foods they can be associated with other molecules, like sugars, proteins or fatty acids. Such associations can result in substantial changes in their properties. Indeed, food xanthophylls are commonly found in the form of esters, above all in many fruits. This modification markedly influences properties such as solubility and susceptibility to oxidation, which in turn can have impacts on relevant aspects that explain their levels in foods and humans, such as their biosynthesis, deposition, stability and bioavailability, among others. The study of the esterification of carotenoids is undoubtedly gaining popularity. In this chapter, nomenclature and structural aspects related to isoprenoids, carotenoids, fatty acids and, finally, carotenoid esters are presented, with references to some physical–chemical properties and their importance at different levels.

Carotenoids are isoprenoid pigments used by pharmaceutical, cosmetic, food and feed industry as antioxidants and colorants. Although traditional sources of carotenoids are fruits, vegetables and chemical synthesis, prospecting for alternative sinks of common and/or unusual carotenoids is important for the development of natural carotenoid industry. In this work, 30 pigmented bacterial strains from Fildes Peninsula in King George Island, Antarctica, were isolated and identified by 16S rRNA gene sequencing and classified in three phyla, Bacteroidetes, Firmicutes and Actinobacteria. After cells extraction, ten different carotenoids were identified based on the chromatographic and spectroscopic characteristic obtained by HPLC-PDA and HPLC-PDA-APCI-MS analyses. Strains assigned to Bacteroidetes affiliated to Flavobacterium, Chryseobacterium and Zobellia genera, presented a pigment profile composed of zeaxanthin, β-cryptoxanthin and β-carotene. Firmicutes strains of Planococcus genus produced a C50 carotenoid, identified as C.p. 450 glucoside. Actinobacteria isolates were mainly assigned to Arthrobacter genus, and few to Salinibacterium and Cryobacterium genera. Arthrobacter strains produced C50 carotenoids such as decaprenoxanthin and its glucosylated derivatives, as well as some C40 carotenoids such as lycopene which is used as synthesis precursors of the C50 carotenoids. Salinibacterium and Cryobacterium genera produced C.p. 450 free form and its glucosylated derivatives. Although most isolates produce carotenoids similar in diversity and quantity than those already reported in the literature, novel sources for C50 carotenoids results from this work. According to their carotenoid content, all isolates could be promising candidates for carotenoids production., This work was supported by “Comisión Sectorial de Investigación Científica” [project CSIC I+D 2014 219], and “Agencia Nacional de Investigación e Innovación” [grant POS_NAC_2014_1102321] and [grant MOV_CA__2017_1_138162].

Volcanic eruptions produce health changes in animals that may be associated with emitted gases and deposited ashes. We evaluated whether the Puyehue–Cordón Caulle volcanic eruption in 2011 produced health changes in the threatened Andean Condor (Vultur gryphus) living in the area most affected by the eruption, northwestern Patagonia. We studied clinical and biochemical parameters of condors examined before and after the eruption. We also examined concentrations of different metals and metalloids in the blood of individuals sampled after the eruption. The most common clinical abnormality associated with the eruptive process was irritating pharyngitis. In condors sampled after the eruption, blood concentrations of albumin, calcium, carotenoids, and total proteins decreased to levels under the reference values reported for this species. We found different chemical elements in the blood of these condors after the eruption, such as arsenic and cadmium, with the potential to produce health impacts. Thus, the health of Andean Condors was affected in different ways by the eruption; remaining in the affected area appears to have been costly. However, in comparison to other animal species, the health impacts were not as strong and were mainly related to food shortages due to the decrease in availability of livestock carcasses linked to the eruption. This suggests that condors dealt relatively well with this massive event. Future research is needed to evaluate if the health changes we found reduce the survival of this species, and if the cost of inhabiting volcanic areas has any ecological or evolutionary influence on the condor’s life history., Las erupciones volcánicas producen alteraciones en la salud de los animales asociadas con los gases emitidos y las cenizas eyectadas. En este trabajo, evaluamos si la erupción del volcán Puyehue–Cordón Caulle ocurrida en 2011 produjo alteraciones en la salud de los Vultur gryphus que habitan el noroeste de la Patagonia, el área más afectada por la erupción. En particular, estudiamos parámetros clínicos y bioquímicos en cóndores capturados antes y después de la erupción. Además, examinamos las concentraciones de diferentes metales y metaloides en la sangre de individuos capturados después de la erupción. La anormalidad clínica más frecuente asociada con el proceso eruptivo fue la faringitis irritativa. Los cóndores muestreados después de la erupción mostraron concentraciones sanguíneas de albúmina, calcio, carotenoides y proteínas totales por debajo de los valores de referencia informados para esta especie. En la sangre de los cóndores muestreados después de la erupción también encontramos diferentes elementos químicos, como arsénico y cadmio, potencialmente dañinos para su salud. Por lo tanto, nuestros resultados sugieren que la permanencia de estos individuos en la zona afectada tuvo costos para su salud. Sin embargo, en comparación con otras especies animales, los impactos no fueron graves y estuvieron relacionados principalmente con la escasez de alimentos debido a la disminución en la disponibilidad de carroñas de ganado vinculada al proceso eruptivo. Futuras investigaciones son necesarias para evaluar si los cambios en la salud encontrados pueden reducir la supervivencia de esta especie, y si el costo de habitar áreas volcánicas tiene alguna influencia ecológica o evolutiva en la historia de vida del Vultur gryphus.

This work was supported by funding from the ‘Ministerio de Ciencia, Innovacion y Universidades’ (Spanish Government, Projects AGL2014-53195R and AGL2017-85368-P).

High irradiances may lead to photooxidative stress in plants, and non-photochemical quenching (NPQ) contributes to protection against excess excitation. One of the NPQ mechanisms, qE, involves thermal dissipation of the light energy captured. Importantly, plants need to tune down qE under light-limiting conditions for efficient utilization of the available quanta. Considering the possible redox control of responses to excess light implying enzymes, such as thioredoxins, we have studied the role of the NADPH thioredoxin reductase C (NTRC). Whereas Arabidopsis thaliana plants lacking NTRC tolerate high light intensities, these plants display drastically elevated qE, have larger trans-thylakoid ΔpH and have 10-fold higher zeaxanthin levels under low and medium light intensities, leading to extremely low linear electron transport rates. To test the impact of the high qE on plant growth, we generated an ntrc-psbs double-knockout mutant, which is devoid of qE. This double mutant grows faster than the ntrc mutant and has a higher chlorophyll content. The photosystem II activity is partially restored in the ntrc-psbs mutant, and linear electron transport rates under low and medium light intensities are twice as high as compared with plants lacking ntrc alone. These data uncover a new role for NTRC in the control of photosynthetic yield.

17 Páginas.-- 5 Figuras, Neurosporaxanthin (NX) is a carboxylic carotenoid produced by some filamentous fungi, including species of the genera Neurospora and Fusarium. NX biosynthetic genes and their regulation have been thoroughly investigated in Fusarium fujikuroi, an industrial fungus used for gibberellin production. In this species, carotenoid-overproducing mutants, affected in the regulatory gene carS, exhibit an upregulated expression of the NX pathway. Based on former data on a stimulatory effect of nitrogen starvation on carotenoid biosynthesis, we developed culture conditions with carS mutants allowing the production of deep-pigmented mycelia. With this method, we obtained samples with ca. 8 mg NX/g dry mass, in turn the highest concentration for this carotenoid described so far. NX-rich extracts obtained from these samples were used in parallel with carS-complemented NX-poor extracts obtained under the same conditions, to check the antioxidant properties of this carotenoid in in vitro assays. NX-rich extracts exhibited higher antioxidant capacity than NX-poor extracts, either when considering their quenching activity against [O2(1Δg)] in organic solvent (singlet oxygen absorption capacity (SOAC) assays) or their scavenging activity against different free radicals in aqueous solution and in liposomes. These results make NX a promising carotenoid as a possible feed or food additive, and encourage further studies on its chemical properties., This work was funded by the Spanish Government (Ministerio de Economía y Competitividad, grants BIO2015-69613-R and RTI2018-101902-B-I00; Ministerio de Ciencia, Innovación y Universidades, grants RTI-2018-095131-B-I00 and AGL2015-70218). Authors belong to the Spanish Carotenoid Network, CaRed, (Ministerio de Economía y Competitividad, grant BIO2015-71703-REDT), and the European Carotenoid Network (EuroCaroten) funded by the European Commission COST Action CA15136.

The ascomycetous fungi Fusarium fujikuroi and Neurospora crassa are widely used as research models in the study of secondary metabolism and photobiology, respectively. Both fungi exhibit a similar carotenoid pathway, for which all the genes and enzymes have been identified. Under standard laboratory conditions, either F. fujikuroi or N. crassa accumulate a mixture of neurosporaxanthin, a carboxylic apocarotenoid acid, and several of its carotene precursors. We formerly described methods for the identification and quantification of neurosporaxanthin. However, the differences in polarity between this acidic xanthophyll and neutral carotenes make their global analysis cumbersome. Here we propose a simple HPLC methodology for the efficient separation of neurosporaxanthin and earlier pathway intermediates in a single HPLC run. This method should be useful to check the abundance of neurosporaxanthin under different experimental conditions and to evaluate the relative proportions of their different carotene precursors. To assess the validity of the method, we have compared the carotenoid profiles in samples of mycelia of F. fujikuroi and conidia of N. crassa, in both cases obtained from surface cultures of a wild strain of each species.

24 Páginas.-- 3 Tablas.-- 1 Figura, Carotenoids, especially β-cryptoxanthin, exert multiple biological activities in the organism. Various processing techniques can improve carotenoid bioavailability in relation to the food matrix. The study objective was to compare the bioavailability of carotenoids from orange juice (OJ) with that from a beverage obtained by alcoholic fermentation of orange juice (FOB). Seven volunteers were recruited for a randomized, controlled, and crossover study. Post-intake plasma carotenoid concentrations were measured by HPLC in the subjects at 0–8 h after their consumption of OJ or FOB. β-Cryptoxanthin and lutein absorption was significantly higher from FOB than from OJ, but no significant difference in zeaxanthin absorption was found. The mean baseline-corrected area under the concentration curve (AUC0-8 h) for β-cryptoxanthin, lutein and zeaxanthin was 24.6-, 1.3- and 4.65-fold larger, respectively, after FOB versus OJ consumption. This fermented orange beverage could be an abundant source of bioavailable carotenoids, and its regular consumption may exert healthy effects., The authors thank the Spanish Government (projects AGL2010-14850/ALI and AGL2014-53195R) and Junta de Andalucía (projects P09-AGR4814M and P08-AGR-03477, and group PAI BIO311) for financial support. DHM is member of CaRed Network funded by MINECO (BIO2015-71703-REDT) and participates in EUROCAROTEN COST Action (CA15136). They are also grateful to Grupo Hespérides Biotech S.L. for supplying the fermented orange beverage.

Carotenoids are lipophilic isoprenoid compounds synthesized by all photosynthetic organisms and some non-photosynthetic prokaryotes and fungi. With some notable exceptions, animals (including humans) do not produce carotenoids de novo but take them in their diets. In photosynthetic systems carotenoids are essential for photoprotection against excess light and contribute to light harvesting, but perhaps they are best known for their properties as natural pigments in the yellow to red range. Carotenoids can be associated to fatty acids, sugars, proteins, or other compounds that can change their physical and chemical properties and influence their biological roles. Furthermore, oxidative cleavage of carotenoids produces smaller molecules such as apocarotenoids, some of which are important pigments and volatile (aroma) compounds. Enzymatic breakage of carotenoids can also produce biologically active molecules in both plants (hormones, retrograde signals) and animals (retinoids). Both carotenoids and their enzymatic cleavage products are associated with other processes positively impacting human health. Carotenoids are widely used in the industry as food ingredients, feed additives, and supplements. This review, contributed by scientists of complementary disciplines related to carotenoid research, covers recent advances and provides a perspective on future directions on the subjects of carotenoid metabolism, biotechnology, and nutritional and health benefits., Authors belong to the Spanish Carotenoid Network (CaRed) funded by the Spanish MINECO (Ministry of Economy, Industry and Competitiveness) grant BIO2015-71703-REDT, the European Carotenoid Network (EuroCaroten) funded by the European Commission COST ActionCA15136, and the Ibero-American Carotenoid Network (IberCarot) funded by the Ibero-American Programme for Science, Technology and Development (CYTED) grant 112RT0445. Research on carotenoids at the authors' individual laboratories was supported by Spanish MINECO grants BIO2014-59092-P to MRC, BIO2015-69613-R to JA and MCL, AGL2013-43522-R to AB, BIO2016-77000-R to LGG, AGL2014-53195-R to DHM, AGL2015-67019-P to AP, BIO2014-54441-P to CZ, and AGL2015-70218 to LZ and MJR; Instituto de Salud Carlos III-FEDER grant PI16/01991 to BOA; Generalitat de Catalunya grants 214SGR-1434 to MRC and AB and 2014SGR-1296 to CZ; and Junta de Andalucia grants P10-CTS-6638 to JA and P12-AGR-1287 to AJMM. We also acknowledge the financial support of the CERCA Programme of the Generalitat de Catalunya and the Severo Ochoa Programme for Centres of Excellence in R&D 2016–2019 to CRAG (SEV-2015-0533).

13 Páginas; 4 Figuras, The ascomycetous fungi Fusarium fujikuroi and Neurospora crassa are widely used as research models in the study of secondary metabolism and photobiology, respectively. Both fungi exhibit a similar carotenoid pathway, for which all the genes and enzymes have been identified. Under standard laboratory conditions, either F. fujikuroi or N. crassa accumulate a mixture of neurosporaxanthin, a carboxylic apocarotenoid acid, and several of its carotene precursors. We formerly described methods for the identification and quantification of neurosporaxanthin. However, the differences in polarity between this acidic xanthophyll and neutral carotenes make their global analysis cumbersome. Here we propose a simple HPLC methodology for the efficient separation of neurosporaxanthin and earlier pathway intermediates in a single HPLC run. This method should be useful to check the abundance of neurosporaxanthin under different experimental conditions and to evaluate the relative proportions of their different carotene precursors. To assess the validity of the method, we have compared the carotenoid profiles in samples of mycelia of F. fujikuroi and conidia of N. crassa, in both cases obtained from surface cultures of a wild strain of each species., We thank the Spanish Government (projects BIO2012-39716, BIO2015-69613-R, AGL2014-53195R, and CaRed Network BIO2015-71703-REDT) and Junta de Andalucía (project CTS-6638) for financial support.

Potato tubers are considered an important source of bioactive compounds, although the concentrations of different phytochemicals are affected by cooking and other processes. The present study was focused on evaluating the effect of boiling on total phenolic, anthocyanin and carotenoid concentrations of ten promising introgressed breeding lines from native potato species (Solanum tuberosum subsp. andigena Hawkes and Solanum stenotomum Juz. & Bukasov) and three heirloom Spanish S. tuberosum L. cultivars (Jesu¿ s, Morada and Zamora). In addition, three commercial cultivars (Vitelotte, Kasta and Monalisa) with different flesh color were used as testers. Breeding lines NK-08/349 and NK-08/362 showed high total phenolic (TP) and total anthocyanin concentrations (TA), whereas breeding lines NK-08/286, NK-08/356 and NK-08/370 and the cultivar Morada were characterized by a high carotenoid concentration (TC). In general, purple cultivars and breeding lines (Vitelotte, Morada, Jesus, Kasta, NK-08/349 and NK-08/362) stood out with showed high phenolic and anthocyanin concentrations. The comparative analysis of raw and boiled tubers showed high losses of phenolics, anthocyanins and carotenoids. TP, TA and TC concentrations in boiled tubers were directly correlated with their corresponding concentration in the raw product. In addition, significant correlations (p 0.05) between TA and TP concentrations, in both raw and boiled tubers, have been observed. The utilization of peeled and diced tubers, with the subsequent surface increase, seems to be a determinant factor explaining why phytochemical losses were so high., This work was supported by funding from the Ministerio de Ciencia e Innovación (Spanish Government, Projects AGL2010-14850/ALI, RTA2011-00018-C03-01 and RFP2012-00006) and the Consejería de Economía, Innovación, Ciencia y Empleo (Junta de Andalucía, Project P08-AGR-03477). DHM and LGG are members of the IBERCAROT Network, funded by CYTED (ref. 112RT0445). Thanks are due to Visitación Ortega Márquez for her technical assistance on phytochemical extractions.

27 páginas, 3 figuras, 2 tablas, The process of in vivo esterification of xanthophylls has proven to be an important part of the post-carotenogenesis metabolism which mediates their accumulation in plants. The biochemical characterization of this process is therefore necessary for obtaining new and improved crop varieties with higher carotenoid contents. This study investigates the impact of postharvest storage conditions on carotenoid composition, with special attention to the esterified pigments (monoesters, diesters and their regioisomers), in durum wheat and tritordeum, a novel cereal with remarkable carotenoid content. For tritordeum grains, the total carotenoid content decreased during the storage period in a clear temperature-dependent manner. On the contrary, carotenoid metabolism in durum wheat was very much dependent on the physiological adaptation of the grains to the imposed conditions. Interestingly, when thermal conditions were more intense (37 °C), a higher carotenoid retention was observed for tritordeum, and was directly related to the de novo esterification of the lutein induced by temperature. The profile of lutein monoester regioisomers was constant during storage, indicating that the regioisomeric selectivity of the XAT enzymes was not altered by temperature. These data can be useful for optimizing the storage conditions of grains favoring a greater contribution of carotenoids from these staple foods., This work was supported by funding from the Ministerio de Ciencia e Innovación (Spanish Government, Projects AGL2010-14850/ALI, AGL2008-03720) and the Consejería de Economía, Innovación, Ciencia y Empleo (Junta de Andalucía, Project P08-AGR-03477). EMO was the recipient of a JAE-Predoctoral grant (CSIC) co-financed by the ESF. DHM and EMO are members of the IBERCAROT Network, funded by CYTED (ref. 112RT0445).

Alcoholic fermentation of fruits has generated novel products with high concentrations of bioactive compounds and moderate alcohol content. The aim of this study was to evaluate the potential effect on cardiovascular risk factors of the regular consumption by healthy humans of a beverage obtained by alcoholic fermentation and pasteurization of orange juice.Thirty healthy volunteers were enrolled in a randomized controlled study. The experimental group (n = 15) drank 500 mL orange beverage (OB) per day for 2 weeks (intervention phase), followed by a 3-week washout phase. Blood samples were collected at baseline (E-T0) and at the end of the intervention (E-T1) and washout (E-T2) phases. Controls (n = 15) did not consume OB during a 2-week period. OB intake significantly increased oxygen radical absorbance capacity (43.9%) and reduced uric acid (-8.9%), catalase (CAT) (-23.2%), thiobarbituric acid reactive substances (TBARS) (-30.2%) and C-reactive protein (-2.1%) (E-T1 vs. E-T0). These effects may represent longer-term benefits, given the decreased uric acid (-8.9%), CAT (-34.6%), TBARS (-48.4%) and oxidized low-density lipoprotein (-23.9%) values recorded after the washout phase (E-T2 vs. E-T0).The regular consumption of OB improved antioxidant status and decreased inflammation state, lipid peroxidation and uric acid levels. Thus OB may protect the cardiovascular system in healthy humans and be considered a novel functional beverage. © 2017 Society of Chemical Industry.

The increase of lutein retention through the food chain is desirable for wheat breeding. Lutein esters are more stable than free lutein during post-harvest storage and two loci on chromosomes 7D and 7Hch are important for esterification. We investigated the effect of temperature during grain filling on carotenoid accumulation and lutein ester profile including fatty acid selectivity (palmitic vs. linoleic) and regioselectivity (esterification at positions 3 vs. 3′). Three different temperature regimes were assayed (controlled, semi-controlled and non-controlled). Lutein esters were more stable than free carotenoids in vivo and the enzymes encoded by chromosomes 7Hch and 7D are complementary. Indeed, they show differential preferences for the fatty acid (palmitic and linoleic, respectively) and regioselectivity (3 and 3′, respectively). Besides, H. chilense has additional genes for esterification. Finally, the increase of temperature favoured the accumulation of lutein esters with linoleic acid and the synthesis of regioisomers at position 3′., This research was funded by Grant AGL2014-53195R, from Ministerio de Economía y Competitividad, Spain (MINECO) including FEDER funding. D.H.-M. is a member of CaRed Network, funded by MINECO, Spain, (BIO-2015-71703-REDT). M.G.M. was recipient of a fellowship from MINECO (BES-2012-055961).

Apple (Malus x domestica Borkh) has always been considered a fruit with low chlorophyll and carotenoid contents; however these pigments contribute also to the external (peel) and internal (flesh) fruit colouration, as well as to the health benefits associated with the regular consumption of this fruit. In the present work we studied the chlorophyll and carotenoid composition of the peel and flesh of thirteen marketed apple varieties presenting different external colouration (green, yellow and red). All the varieties were characterised by a common pigment profile comprised by chlorophylls a and b, lutein, violaxanthin, neoxanthin, β-carotene, and esterified carotenoids (mainly violaxanthin and neoxanthin) as major compounds. Total pigment content was always higher in the peel (58.72-1510.77 μg/g dry wt) than in the flesh (14.80-71.57 μg/g dry wt) for each particular cultivar. In general, the green cultivars showed the highest pigment content both in the peel and the flesh, which were followed in decreasing order by the peel of some red-skinned cultivars and the flesh of the yellow ones. In most cultivars, a characteristic chloroplastic pigment profile was found in the peel, with lutein as the main free carotenoid while a chromoplastic profile was usually observed in the flesh, with a prevalence of violaxanthin and neoxanthin and their corresponding acyl esters (mono- and diesters). A correlation between the carotenoid content and the amount of esterified xanthophylls was observed, in particular in the flesh, which reinforce the putative role of the esterification process in the accumulation of these lipophilic compounds within the chromoplasts.

A high carotenoid content is important for the production of pasta from durum wheat (Triticum durum Desf.) and yellow alkaline noodle from common wheat (T. aestivum L.). Carotenoid esters are more stable than free carotenoid during storage and processing, and thus they allow a higher retention through the food chain. Chromosome 7D carries gene(s) for lutein esterification. The aim of this study was the physical mapping of the gene(s) for lutein esterification on chromosome 7D and the identification of candidate genes for this trait. We developed crosses between a set of deletion lines for chromosome 7D in Chinese Spring (CS) background and the CS–Hordeum chilense substitution line CS(7D)7Hch. The F2 progeny derived from the deletion line 7DS4 produced a lower amount of lutein esters, which indicates that the main gene for lutein esterification is in the region of chromosome 7D lacking in 7DS4. Other gene(s) are contributing to lutein esterification because small amounts of lutein esters are produced in 7DS4. Genotyping by DArTSeq revealed that 7DS4 lacks a 127.7 Mb region of 7DS. A set of 10 candidate genes for lutein esterification was identified by using the wheat reference genome sequence along with the Wheat Expression Browser. This region contains the Lute locus previously identified in a different genetic background. Four genes with acyltransferase or GDSL esterase/lipase activity were identified in the vicinity of Lute. Our results indicate that the gene TraesCS7D01G094000 is a likely candidate for Lute but the gene TraesCS7D01G093200 cannot be ruled out. The candidate genes reported in this work are worthy for further investigation., This research was funded by Grants AGL2014-53195-R from Ministerio de Economía y Competitividad and AGL2017-85368-P from Ministerio de Ciencia, Innovación y Universidades. Both projects are cofounded by FEDER. DHM is member of CaRed Network funded by Ministerio de Economía y Competitividad (BIO2015-71703-REDT and BIO2017-90877-REDT) and participates in EUROCAROTEN COST Action (CA15136). CMA and SGA are members of FiRCMe Network, funded by Ministerio de Economía y Competitividad (AGL2016-81855-REDT).

The intake of bioactive compounds and moderate alcohol decreases the risk of cardiovascular diseases. These effects could be joined in a beverage created by a controlled alcoholic fermentation of orange juice. The influence of controlled alcoholic fermentation on the bioactive compound profile of orange juice has not been previously evaluated, and this is the purpose of the present study. Total and individual flavanones and carotenoids significantly increased throughout the fermentation. The reason for this was an enhanced extraction of these compounds from the pulp. Besides, the potential bioavailability of flavanones increased due to a higher content of hesperetin-7-O-glucoside (2-fold higher at the end of the fermentation process). Ascorbic acid did not undergo a significant change, and only total phenolics decreased. Antioxidant capacity was also evaluated. TEAC and FRAP values remained constant throughout the process. However, ORAC and DPPH values significantly increased. Correlation analysis concluded that the increase in ORAC and DPPH values could be due to enhancement of flavanones.