Hacia el uso de carotenoides incoloros como ingredientes funcionales de alimentos.

Durante el periodo de la tesis se llevaron a cabo estudios in vitro para evaluar la bioaccesibilidad del fitoeno (PT) y del fitoflueno (PTF) en varias matrices alimentarias y determinar su eficiencia en la micelización y ensayos celulares para estudiar la captación y el eflujo intestinal de los mismos. Con estos estudios se trató de evaluar varios aspectos de estos carotenoides que son útiles para su uso como ingredientes funcionales de alimentos y/o productos relacionados y de identificar las posibles características diferenciales entre estos carotenoides y otros carotenoides biodisponibles de la dieta. En primer lugar, con el fin de obtener información de base para el desarrollo de la tesis, se realizó un estudio bibliográfico que dio lugar a un capítulo de libro y dos revisiones que se han incluido como parte de la introducción de la presente tesis. En los primeros estudios experimentales se evaluó la bioaccesibilidad del PT y del PTF de varias matrices alimentarias ricas en estos carotenoides mediante digestiones in vitro gastrointestinales. Las matrices evaluadas fueron varios zumos comerciales de frutas y de hortalizas, un zumo de naranja de la variedad Pinalate (mutante rica en carotenoides incoloros), un tomate fresco común, un tomate fresco tipo cherry y dos productos de tomate en polvo. En estos estudios se observó cómo, con carácter general e independientemente de la matriz analizada, la bioaccesibilidad del PT era algo superior a la del PTF y cómo las bioaccesibilidades de ambos eran superiores a la de otros carotenoides biodisponibles presentes en las matrices, siendo particularmente superiores a la del licopeno, otro caroteno lineal. Por ejemplo, en el zumo de tomate, una de las frutas con mayor contenido en estos carotenoides, la bioaccesibilidad del PTF y del PT (50 y 62%, respectivamente) fue aproximadamente 3 y 4 veces superior a la del licopeno, respectivamente. Dado que los diferentes isómeros geométricos del PT y del PTF mostraron, al contrario que los del resto de los carotenoides analizados, bioaccesibilidades similares entre sí, la mayor proporción de isómeros cis de los mismos presentes en la mayoría de las matrices en relación a la del resto de los carotenoides analizados podría no ser la causa principal de la mayor bioaccesibilidad de los mismos. Con los resultados de los estudios in vitro sobre la micelización de carotenoides se concluyó que la mayor bioaccesibilidad del PT y del PTF podría ser consecuencia de su mayor capacidad para incorporarse en las micelas mixtas durante la digestión. A pesar de que generalmente se considera que la bioaccesibilidad de un carotenoide es mayor cuanto mayor es su polaridad, la elevada capacidad de micelización y bioaccesibilidad del PT y del PTF parecían deberse, principalmente, a su mayor flexibilidad y a su característica forma molecular, que son consecuencia de su menor número de dobles enlaces conjugados en comparación con el del resto de carotenoides comunes. En el estudio con muestras de tomate en polvo se pudo comprobar como la adición de aceite de girasol conducía a un incremento de la bioaccesibilidad del PT y del PTF, ya de por sí alta. Así, de forma aproximada, se observó como la bioaccesibilidad del PT se triplicó y la del PTF se cuadriplicó tras la adición de un 5% (volumen/peso fresco) de aceite. Por otra parte, se analizó el efecto de varios tratamientos térmicos en un zumo de naranja fresco de la variedad Pinalate en la concentración y en la bioaccesibilidad de los carotenoides principales presentes en la matriz y en el color del zumo. Se observó cómo todos los tratamientos estudiados producían una disminución significativa de la concentración de los carotenoides incoloros (p < 0,05). Se llegó a la conclusión de que ciertos tratamientos térmicos producían un aumento significativo (p < 0,05) de la bioaccesibilidad de los carotenoides incoloros como consecuencia de la degradación del material celular (observado mediante microscopía) y de la disminución del tamaño de partícula (analizado mediante el estudio de la distribución del tamaño de partícula). El zumo pasteurizado fue el que generó mayores diferencias con el zumo fresco en relación a la degradación de las estructuras celulares, el tamaño de partícula, el color, la concentración de carotenoides y la bioaccesibilidad de los mismos. Entre los zumos de frutas y de hortalizas comerciales analizados, el zumo de tomate fue el que tuvo una mayor concentración en carotenoides incoloros y el que proporcionó un mayor contenido bioaccesible de estos carotenoides. Así, el consumo de 250 mL de zumo de tomate, al que se le adicionó un 33% de crema para café como fuente de grasa, resultaría en 5,8 mg de PT y 2,2 mg de PTF ingeridos y proporcionaría 3,6 mg de PT y 1,1 mg de PTF bioaccesibles. A continuación, se procedió a estudiar el proceso de captación y eflujo intestinal en células Caco-2. La eficiencia de captación intestinal del PTF y del PT fue similar a la de la luteína y superior a la del licopeno (29, 21, 26% y no detectable, respectivamente). Estudios de competencia por la captación intestinal mostraron que la presencia de PTF, β-caroteno o luteína micelar en la cámara apical disminuía significativamente la captación del PT micelar (p < 0,001), mientras que solo la luteína micelar, entre los carotenoides analizados, producía una disminución significativa de la captación de PTF micelar (p < 0,001). Mediante el uso de células Caco-2 y HEK se confirmó que la proteína SR-BI estaba implicada en la captación intestinal del PT y del PTF. Se observó que parte del PT y del PTF micelar captado, en particular aproximadamente el 14%, podía sufrir eflujo de vuelta a la cámara apical. Este estudio parece ser el primero en el que se obtienen evidencias experimentales que indican que los carotenoides pueden sufrir eflujo intestinal.During the period of this doctoral thesis, in vitro studies were carried out to evaluate the bioaccessibility of phytoene (PT) and phytofluene (PTF) in several food matrices and to determine their micellization efficiencies and cellular experiments were performed to study their intestinal uptake and efflux. With these studies we tried to evaluate several aspects of these carotenoids that are useful for their use as functional ingredients of food and / or related products and to identify the possible differential characteristics between these carotenoids and other bioavailable carotenoids of the diet. The aims of these studies were try to evaluate several aspects of these carotenoids that are useful for their use as functional ingredients of food and/or related products and to identify the possible differential characteristics between these carotenoids and other bioavailable carotenoids of the diet. First of all, a bibliographical study was conducted in order to obtain basic information for the development of the thesis. One book chapter and two reviews, which have been included as part of the introduction of the present thesis, were the fruits of this labour. In the first experimental studies, the bioaccessibility of PT and PTF in several foods rich in them was evaluated by in vitro gastrointestinal digestions. The matrices evaluated were several commercial juices of fruit and vegetables, an orange juice of the Pinalate variety (a mutant rich in colourless carotenoids), a fresh common tomato, a fresh cherry tomato and two tomato powder products. In these studies, it was observed that, in general and independently of the analysed matrix, the bioaccessibility of PT was slightly higher than that of PTF and that the bioaccessibility of both was superior to that of other bioavailable carotenoids present in the matrices, being particularly superior to that of lycopene, another linear carotene. For example, in the tomato juice, one of the fruits with the highest content of these carotenoids, the bioaccessibility of PTF and PT (50 and 62%, respectively) was approximately 3 and 4 times higher than that of lycopene, respectively. Given that the different geometrical isomers of PT and PTF showed, unlike the rest of the carotenoids analysed, similar bioaccessibilities to each other, the higher proportion of cis isomers of PT and PTF present in most of the matrices in relation to that of the rest of the analysed carotenoids may not be the main cause of their greater bioaccessibility. The in vitro micellization studies indicated that the higher bioaccessibility of PT and PTF could be a consequence of its greater capacity to be incorporated in mixed micelles during digestion. Although it is widely accepted that the greater the polarity of a carotenoid, the greater its bioaccessibility, the high micellization efficiency and bioaccessibility of PT and PTF seemed to be due, mainly, to its greater flexibility and its characteristic molecular shape, which are a consequence of its lower number of conjugated double bonds compared to that of the rest of common carotenoids. In the study with the tomato powder samples, it was observed that the addition of sunflower oil increased the bioaccessibility of PT and PTF, which was already high. Approximately, the bioaccessibility of PT tripled and that of PTF quadrupled after the addition of 5% (volume / fresh weight) of oil. On the other hand, the effect of several thermal treatments in a fresh juice of the Pinalate orange on the concentration and bioaccessibility of the main carotenoids present in the matrix and in the colour of the juice was analysed. It was noted that all the treatments studied produced a significant decrease in the concentration of colourless carotenoids (p < 0.05). It was concluded that certain thermal treatments produced a significant increase (p < 0.05) of the bioaccessibility of the colourless carotenoids as a consequence of the degradation of cellular material (examined by microscopy) and the decrease in particle size (analysed by studying the particle size distribution). The pasteurization was the process that gave rise to the juice with greater differences with respect to the fresh juice in relation to the degradation of the cellular structures, the particle size, the colour, and the concentration and bioaccessibility of carotenoids. Among the commercial juices analysed, the tomato juice was the sample with the highest concentration of colourless carotenoids and the juice that provided the highest bioaccessible content of them. Thus, the consumption of 250 mL of the tomato juice, to which 33% of coffee cream was added as a source of fat, would result in an intake of 5.8 and 2.2 mg of PT and PTF, respectively, and would provide 3.6 and 1.1 mg of bioaccessible PT and PTF, respectively. Subsequently, the intestinal uptake and efflux were studied by in vitro experiments using Caco-2 cells. The uptake efficiency of PTF and PT were similar to that of lutein and superior to that of lycopene (29, 21, 26% and not detectable, respectively). Competition studies for intestinal uptake showed that the presence of micellar PTF, β-carotene or lutein in the apical chamber significantly decreased the micellar PT uptake (p <0.001), whereas only micellar lutein, among the carotenoids analysed, produced a significant decrease in micellar PTF uptake (p < 0.001). By using Caco-2 and HEK cells it was confirmed that SR-BI protein is involved in the intestinal uptake of both PT and PTF. It was observed that part of the micellar PT and PTF taken up by Caco-2 cells, in particular about 14%, could undergo efflux back into the apical chamber. This seems to be the first study showing that carotenoids may suffer intestinal efflux from experimental evidences.Premio Extraordinario de Doctorado U

Microalgae, Seaweeds and Aquatic Bacteria, Archaea, and Yeasts: Sources of Carotenoids with Potential Antioxidant and Anti-Inflammatory Health-Promoting Actions in the Sustainability Era

Carotenoids are a large group of health-promoting compounds used in many industrial sectors, such as foods, feeds, pharmaceuticals, cosmetics, nutraceuticals, and colorants. Considering the global population growth and environmental challenges, it is essential to find new sustainable sources of carotenoids beyond those obtained from agriculture. This review focuses on the potential use of marine archaea, bacteria, algae, and yeast as biological factories of carotenoids. A wide variety of carotenoids, including novel ones, were identified in these organisms. The role of carotenoids in marine organisms and their potential health-promoting actions have also been discussed. Marine organisms have a great capacity to synthesize a wide variety of carotenoids, which can be obtained in a renewable manner without depleting natural resources. Thus, it is concluded that they represent a key sustainable source of carotenoids that could help Europe achieve its Green Deal and Recovery Plan. Additionally, the lack of standards, clinical studies, and toxicity analysis reduces the use of marine organisms as sources of traditional and novel carotenoids. Therefore, further research on the processing of marine organisms, the biosynthetic pathways, extraction procedures, and examination of their content is needed to increase carotenoid productivity, document their safety, and decrease costs for their industrial implementation.C.R. and J.M. contributions were financed by The Research Council of Norway (326803 SUSKELPFOOD; 319577 SAFER-IMTA, 294946 SBP-N) and Møreforsking AS. P.M.-B. and A.J.M.-M. acknowledge Grant PID2019-110438RB-C21 (NEWCARFOODS), funded by MCIN/AEI/10.13039/501100011033. P.G.-V., A.L.-V. and R.L. acknowledge FITORIBOPEP-P20_00728. PROY. I + D + I JUNTA ANDALUCIA 2020, AGL2019-110438RB-C22-AEI/FEDER, Fertinagro Biotech Foundation (Cátedra Fertinagro Biotech de la Universidad de Huelva). P.M.-B. was supported by a posdoc fellowship from Consejería de Transformación Económica, Industria, Conocimiento y Universidades de la Junta de Andalucía (PAIDI 2020). P.G.-V. and A.L.-V. wants to thank also Next Generation European Funds and the Ministry of Universities of Spain for funding the Recualificación del Profesorado Universitario system. V.S. and E.V. acknowledge CSIC I + D 2018 250 UdelaR, ANII POS NAC 2014 1 102321, and Ph.D. grant CAP UdelaR 2017-2019

Extraction of carotenoids from cantaloupe waste and determination of its mineral composition

The carotenoid and mineral levels as well as the in vitro antioxidant capacity, using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, of waste from cantaloupe was assessed. Then the matrix was subjected to ultrasound-assisted extraction (UAE) and response surface methodology (RSM) was used for the optimization of the extraction of carotenoids. The effect of the extraction procedure on the microstructure of the powder was assessed by scanning electron microscopy (SEM) analysis. The major carotenoids identified were lutein (63.24 ± 0.73 μg βCE/g dw) and β-carotene (56.43 ± 0.11 μg βCE/g dw). Several mineral elements (K, Na, P, Mg, Ca, Fe, Cu, Mn and Zn) were identified, potassium being the major one.The extract exhibited in vitro antioxidant activity (IC50 = 7.33 ± 0.22 μg/mL). The RSM results showed that an amplitude of 100%, extraction time of 10 min, hexane percentage of 80% in hexane/acetone solvent, and solvent-to-solid ratio of 55 mL/g were the optimal conditions for the extraction of carotenoids. Under these conditions, the carotenoid content of the extract was 124.61 ± 3.82 μg/g.The microscopic analysis revealed the effectiveness of the ultrasound treatment that results in noticeable physical changes, like microscopic perforations and breakages

Isoprenoids composition and colour to differentiate virgin olive oils from a specific mill

The objective of this work was verify if, based on the concentration of some key isoprenoids and the colour, olive oils from a specific mill could be differentiated from those from other mills by linear discriminant analysis. The isoprenoids studied were β-carotene, lutein, α-tocopherol, β-sitosterol, pheophytin a and squalene, which are isoprenoids of interest in food science and nutrition. 88 % of correct classification was obtained by linear discriminant analysis and, therefore, a good differentiation was achieved. This fact can reveal the impact of the conditions of elaboration of each particular mill on the isoprenoid content of the olive oil and therefore on its colour, which is a characteristic of foods with a high effect on consumers

Assessment of Food Sources and the Intake of the Colourless Carotenoids Phytoene and Phytofluene in Spain.

Phytoene (PT) and phytofluene (PTF), colorless carotenoids, have largely been ignored in food science studies, food technology, and nutrition. However, they are present in commonly consumed foods and may have health-promotion effects and possible uses as cosmetics. The goal of this study is to assess the most important food sources of PT and PTF and their dietary intakes in a representative sample of the adult Spanish population. A total of 62 food samples were analyzed (58 fruit and vegetables; seven items with different varieties/color) and carotenoid data of four foods (three fruits and one processed food) were compiled. PT concentration was higher than that of PTF in all the foods analyzed. The highest PT content was found in carrot, apricot, commercial tomato juice, and orange (7.3, 2.8, 2.0, and 1.1 mg/100 g, respectively). The highest PTF level was detected in carrots, commercial tomato sauce and canned tomato, apricot, and orange juice (1.7, 1.2, 1.0, 0.6, and 0.04 mg/100 g, respectively). The daily intakes of PT and PTF were 1.89 and 0.47 mg/person/day, respectively. The major contributors to the dietary intake of PT (98%) and PTF (73%) were: carrot, tomato, orange/orange juice, apricot, and watermelon. PT and PTF are mainly supplied by vegetables (81% and 69%, respectively). Considering the color of the edible part of the foods analyzed (fruit, vegetables, sauces, and beverages), the major contributor to the daily intake of PT and PTF (about 98%) were of red/orange color.post-print1287 K

Phytoene and Phytoene-Rich Microalgae Extracts Extend Lifespan in C. elegans and Protect against Amyloid-β Toxicity in an Alzheimer’s Disease Model

Phytoene is a colourless carotenoid widely available from dietary sources and a precursor for the synthesis of other carotenoids. Although present at high concentrations across different tissues, phytoene is largely viewed as not having physiological activity. Here, we utilize the model organism C. elegans to show that phytoene is bioactive and has anti-ageing properties. Supplementation with phytoene protects against oxidative damage and amyloid-β42 proteotoxicity (a major pathology of Alzheimer’s disease), and extends lifespan. We also examine extracts from two microalgae, Chlorella sorokiniana and Dunaliella bardawil. We show that the extracts contain high levels of phytoene, and find that these phytoene-rich extracts have protective effects similar to pure phytoene. Our findings show that phytoene is a bioactive molecule with positive effects on ageing and longevity. Our work also suggests that phytoene-rich microalgae extracts can utilized to produce foods or supplements that promote healthy ageing and prevent the development of chronic age-related diseases

Effect of regulated deficit irrigation on commercial quality parameters, carotenoids, phenolics and sugars of the black cherry tomato (Solanum lycopersicum L.) ʽSunchocolaʼ

In this preliminary study, the effect of regulated deficit irrigation (RDI) on the commercial quality (size, weight, soluble solids, firmness and colour), content of carotenoids, phenolics and sugars of black tomato (Solanum lycopersicum L.) ʽSunchocolaʼ was studied. Two water irrigation treatments were applied: regulated deficit irrigation (RDI) and control with 82,7 and 398 mm of water supplied, respectively. Tomato of the first cluster harvested at three stages of maturity were studied. The size and weight of the tomato did not present significant differences regarding the RDI and maturity. In both groups the concentration of carotenoids and phenolics increased with the degree of maturity (on average 57 % and 8 % respectively). On the other hand, in most cases, the content of carotenoids, phenolics and sugars showed significant differences between irrigation treatments (p < 0.1). In conclusion, with the application of the RDI, it was possible to maintain the size and weight and increase the carotenoid levels of the fruits.Secretaría Nacional de Educación Superior, Ciencia, Tecnología e Innovacion (SENESCYT)Ministerio de Economía y Competitividad AGL2012-37610Ministerio de Economía y Competitividad BIO2015-71703- RED

Carotenoides en agroalimentación y salud

Los carotenoides son compuestos especiales; si bien es común referirse a ellos como pigmentos, lo cierto es que son compuestos de gran versatilidad e importancia en la naturaleza. Más específicamente, son de gran interés en agroalimentación y salud. Así, por ejemplo, son pigmentos naturales y por lo tanto tienen un importante papel en la elección de alimentos por parte de los consumidores. Asimismo, algunos de ellos son precursores de la vitamina A. Sin embargo, que cada vez exista más interés en los carotenoides en este contexto se debe en gran parte a muchos estudios de distinta naturaleza que indican que pueden proporcionar beneficios para la salud. Su interés en alimentación funcional es por lo tanto indudable. En este libro se refleja la experiencia en carotenoides de un gran número de profesionales de la región iberoamericana. En conjunto, se ofrece una visión general de la investigación sobre estos compuestos en agroalimentación y salud. Los autores son miembros de la red IBERCAROT (http://carotenoides.us.es), que tiene entre sus objetivos conformar una red estable y funcional de profesionales que aúnen esfuerzos en pos de identificar nuevas fuentes de carotenoides de interés nutricional, mejorar su producción y aumentar el valor de los productos que los contengan

Investigación en color y calidad de alimentos

En esta ponencia se pretende presentar a los alumnos de la Escuela Politécnica Superior, las líneas de investigación del grupo Color y Calidad de Alimentos de la Universidad de Sevilla (AGR225, PAIDI) y las posibilidades de realizar Trabajos postdoctorales o continuar las salidas profesionales en el campo de industrias alimentarias.This paper tries to introduce to the student of Polytechnic School, the research lines of Color and Food Quality Group of the University of Seville (AGR225, PAIDI), and the possibilities of carrying out postdoctoral works or continuing the career in the field of food industries