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An optimized PLE method was applied to several truffle species using three different solvent mixtures to obtain bioactive enriched fractions. The pressurized water extracts contained mainly (1 → 3),(1 → 6)-β-D-glucans, chitins, and heteropolymers with galactose and mannose in their structures. The ethanol extracts included fatty acids and fungal sterols and others such as brassicasterol and stigmasterol, depending on the species. They also showed a different fatty acid lipid profile depending on the solvent utilized and species considered. Ethanol:water extracts showed interesting lipids and many phenolic compounds; however, no synergic extraction of compounds was noticed. Some of the truffle extracts were able to inhibit enzymes related to type 2 diabetes; pressurized water extracts mainly inhibited the α-amylase enzyme, while ethanolic extracts were more able to inhibit α-glucosidase. Tuber brumale var. moschatum and T. aestivum var. uncinatum extracts showed an IC50 of 29.22 mg/mL towards α-amylase and 7.93 mg/mL towards α-glucosidase. Thus, use of the PLE method allows o bioactive enriched fractions to be obtained from truffles with antidiabetic properties.

Shiitake mushrooms (Lentinula edodes) contain compounds with hypocholesterolemic and immune-modulatory activities such as ergosterol and chitin that can be partially transformed into vitamin D and chitosan to yield extracts with enhanced bioactivities. A method was optimized to increase vitamin D2 levels by irradiating a non-soluble fraction obtained from shiitake mushrooms during 1 h with UV-light (254 nm) at 50 °C in methanol. After 1 h, almost all of the ergosterol was transformed into vitamin D2. The conversion ratio can be simply adjusted by reducing the irradiation time. A deacetylation treatment with 50% NaOH for 24 h at 95 °C was needed to generate chitosan and other water-soluble lower molecular weight derivatives from chitin. To obtain an extract enriched in both compounds, the defined methods can be combined, but the extracts should be firstly deacetylated and later irradiated. In this order, an extract is obtained containing vitamin D2 (4.65 mg/g) and chitosan (2.83%).

A supercritical fluid extraction methodology was used to extract flavoring and bioactive compounds from truffles. Some parameters such as CO2 flow rate (1–3 mg/mL), extraction time (15–90 min) and different trapping food matrices (grape seed oil, gelatin, agar agar and water) were optimized using response surface methodology to enhance extraction and trapping yields. The optimal conditions (2.27 mg/mL CO2 flow rate, 82.5 min when using 40 °C and 30 MPa, with 1 mL grape seed oil as trapping matrix) obtained with Tuber melanosporum were applied to three different truffle species: Terfezia claveryi, Tuber aestivum and Tuber indicum. A total of 32 metabolites were profiled in the extracts using ultra-high-performance supercritical fluid chromatography coupled to quadrupole time-of-flight mass spectrometry. Compounds such as brassicasterol ergosta-7,22-dienol, oleic and linoleic acid were found at similar amounts in all the extracts but other molecules (e.g. fungal sterols) showed a particular distribution depending on the specie studied and whether a trapping matrix was used at the SFE outlet.

Although genomes from many edible mushrooms are sequenced, studies on fungal micro RNAs (miRNAs) are scarce. Most of the bioinformatic tools are designed for plants or animals, but the processing and expression of fungal miRNAs share similarities and differences with both kingdoms. Moreover, since mushroom species such as Agaricus bisporus (A. bisporus, white button mushroom) are frequently consumed as food, controversial discussions are still evaluating whether their miRNAs might or might not be assimilated, perhaps within extracellular vesicles (i.e., exosomes). Therefore, the A. bisporus RNA-seq was studied in order to identify potential de novo miRNA-like small RNAs (milRNAs) that might allow their later detection in diet. Results pointed to 1 already known and 37 de novo milRNAs. Three milRNAs were selected for RT-qPCR experiments. Precursors and mature milRNAs were found in the edible parts (caps and stipes), validating the predictions carried out in silico. When their potential gene targets were investigated, results pointed that most were involved in primary and secondary metabolic regulation. However, when the human transcriptome is used as the target, the results suggest that they might interfere with important biological processes related with cancer, infection and neurodegenerative diseases.

Truffles are a well-known worldwide product mainly appreciated by their unique aroma, which is composed by more than 50 volatile compounds. However, to this day, no one has accomplished to find the aromatic key that evokes the real aroma of truffles for its use as food flavoring. Among them, black truffle was selected for extraction with supercritical fluids using CO2 as solvent recovering natural truffle aroma fraction. To achieve the optimal extraction ratio, time, pressure and grapeseed oil addition to the separators were evaluated. Aroma from black truffle powder, extracts obtained, and residual cakes fractions were characterized by headspace gas chromatography-spectrometry and olfactometry techniques. The results indicated that optimal extraction conditions were 30 MPa for 3 h. Also, grapeseed oil addition enhanced trapping some key truffle aromatic compounds as 2,3-butanodione, 2-methyl-1-butanol, octanal and dimethyl disulphide. Olfactometry study showed the aromatic profile of the extracts indicating the molecules ethyl pentanoate (fruity), 1-hexen-3-one (metallic) and ethyl hexanoate (fruity) as the main compounds of extracts samples. For the first time, a natural truffle aroma has been obtained using low-value truffles. After aromatic extraction, carbohydrates, proteins, and phenolic compounds were analysed within the residues, showing a potential source of bioactive compounds.

The use of acid oils and fatty acid distillates in poultry feed formulations is limited due to controversial productive results that has been partially related to a lower digestibility compared with their corresponding crude oils. In vitro digestion and bioaccessibility models mimicking fat intestinal digestion will provide valuable information to understand differences in dietary fat digestibility observed in in vivo trials. The hydrolysis processes that occur during intestinal digestion of soybean acid oil (SA: unsaturated), palm fatty acid distillate (PFAD: saturated), and their corresponding crude oils, as well as their potential bioaccessibility were studied using an in vitro digestion model. The release of lipid-class products (Triacylglycerol(s), TAG; Diacylglycerol(s), DAG; Monoacylglycerol(s), MAG; and Free fatty acid(s), FFA) was followed at 10, 30, and 60 min of digestion. In all oils studied, the largest hydrolysis rate occurred within the first 10 min. At the end, the percentages of generated lipid products were in the range of expected values according to the pancreatic lipase activity. After digestion, dietary mixed micelles (DMM) and the non-bioaccessible precipitated phase (PP) were isolated to analyze their lipid-class composition. Unsaturated oils showed higher DMM fraction than saturated ones (P = 0.001). Higher PP (P < 0.001) were generated in acid oils. Results suggested that hydrolysis did not limit oil digestion and that the oil FFA levels influenced their bioaccessibility less than the saturation degree.

Tartrazine (E102) is a synthetic food coloring, which belongs to the class of mono azo dyes and is known to cause numerous health problems. The current research aimed to evaluate the effect of this food dye on the enzymatic activity of amylase, lipase and proteases after a subchronic ingestion in Swiss mice. Additionally, an in vitro digestion model was used to highlight the relationship between the probable toxicity of tartrazine and the nature of the food ingested. The results show that there were no adverse effects of tartrazine on the body weight gain, and on amylase or lipase activities. However, in the high dose of tartrazine (0.05%) group, a significant decrease in trypsin and chymotrypsin enzymatic activities were observed. Regarding the in vitro digestion model, our findings show that there were no changes in the trypsin and chymotrypsin enzymatic activities either using 7.5 or 75 mg of tartrazine mixed with rice, butter or milk. We conclude that excessive consumption of tartrazine appears to alter the enzymatic activity of proteases in vivo which may have deleterious consequences on digestion. Even thought the dose close to the acceptable daily intake does not affect those activities, a strict control of tartrazine dose in high-consumption foods especially among children is an indispensable task.

Eritadenine is a hypocholesterolemic compound that is found in several mushroom species such as Lentinula edodes, Marasmius oreades, and Amanita caesarea (1.4, 0.7 and 0.6 mg per g dry weight, respectively). It was synthesized during all developmental stages, being present in higher concentrations in the skin of shiitake fruiting bodies. When subjected to traditional cooking, grilling followed by frying were more adequate methodologies than boiling or microwaving to maintain its levels. Modern culinary processes such as texturization (with agar-agar) and spherification (with alginate) also interfered with its release. Grilling and gelling using gelatin enhanced eritadenine's bioaccessibility in an in vitro digestion model. An animal model (where male and female rats were administered 21 and 10 mg per kg animal per day of eritadenine) indicated that intake of the compound was safe under these concentrations; it reached the liver and reduced the atherogenic index (TC/HDL) in rat sera. Thus, it might be used to design a functional food.

Three extraction methods were sequentially combined to obtain fractions from Lentinula edodes (shiitake mushrooms) containing bioactive compounds against cardiovascular diseases (CVDs). Fruiting bodies were first extracted with plain water, obtained residue was then submitted to supercritical fluid extraction (SFE) and remaining residue submitted to hot water extraction. Sequential design allowed reutilization of the nonextracted material as raw material for the successive extractions increasing extraction yields and separating interesting compounds. Obtained fractions contained different amounts of ß-glucans, chitins, eritadenine, lenthionine, ergosterol, proteins/peptides and phenolic compounds conferring them different bioactivities. Water soluble fractions showed high antioxidant activities (ABTS+• and DPPH• scavenging capacity and reducing power), they were also able to inhibit one of the main enzymes involved in hypertension (angiotensin-I converting enzyme) and the key enzyme of cholesterol metabolism (3-hydroxy-3-methylglutaryl coenzyme A reductase). The latter inhibitory activity was also noticed in SFE extracts although ergosterol and other lipid-like molecules were isolated. Dietary fibers were separated in the third extraction. Therefore, with this sequential extraction procedure bioactive compounds against CVDs can be selectively separated from a single batch of shiitake powder.

La seta shiitake (Lentinula edodes) contiene moléculas con importantes actividades beneficiosas para la salud humana (hipocolesterolémica, antioxidante, inmunomodulatoria, anticancerígena, antihipertensiva, etc.). La extracción de compuestos lipofílicos de hongos comestibles utilizando CO2 supercrítico ya ha sido empleada para la obtención de extractos ricos en compuestos hipocolesterolémicos, como el ergosterol y sus derivados. Además, los residuos resultantes de la extracción supercrítica también pueden ser aprovechados por su contenido en compuestos fenólicos y polisacáridos (ß-glucanos) con probada actividad antioxidante in vitro. En el presente trabajo se utilizó una planta de extracción a escala piloto para la obtención de diferentes fracciones bioactivas a partir de polvo de shiitake. Partiendo de unas condiciones fijas de presión y temperatura (225 bar y 40 ºC), se llevó a cabo un estudio cinético del rendimiento de extracción (1-5 h) y se recogieron tres fracciones de diferente naturaleza: el extracto precipitado a 100 bar (1), el extracto precipitado a 60 bar (2) y el residuo resultante de la extracción. Los extractos (1) y (2) presentaron concentraciones de ergosterol y sus derivados significativamente superiores al polvo de partida y al residuo. Por el contrario, el residuo presentó una mayor concentración en compuestos fenólicos que el polvo de partida y el extracto, así como una mayor capacidad antioxidante (dependiendo del método de determinación).

Black truffle (Tuber melanosporum) is considered a gourmet product due to its sensory properties, particularly because of its flavour. However, they also showed an interesting nutritional composition i.e. they are low fat, high dietary fibre food and contain many biological compounds such as antioxidants, hypocholesterolemic sterols, etc. Ergosterol (ergosta-5,7,22-trienol) and brassicasterol (ergosta-5,22-dienol) are the two major fungal sterols of T. melanosporum hyphae representing respectively almost 60 and 40% of total sterols. Ergosterol is a vitamin D2 precursor since it is transformed into previtamin D2 which turns into ergocelciferol (vitamin D2) by ultraviolet light (UV) exposure. Vitamin D-enriched mushroom can also be achieved by fruiting bodies irradiation however, to our knowledge no report has been published studying the irradiation of truffles carpophores to increase their vitamin D2 levels. Therefore, in this work, the effect of UV irradiation at two wavelengths on black truffles was studied to investigate whether they were able to transform ergosterol into ergocalciferol. Thus, fresh T. melanosporum ascocarps were irradiated with UV-light (254 or 312 nm) for 30, 60 and 90 min. Ergosterol levels were determined by GC–MS-FID2 and vitamin D2 quantification was carried out using an HPLC method. T. melanosporum ascocarps contained no ergocalciferol since they are usually buried in the woods but showed 0.8-1.8 mg/g ergosterol and 0.5-0.7 mg/g brasicasterol. They also contained other sterols such as ergosta7,22-dienol and 9,19ciclolanost-7-en-3-ol. Truffle UV-irradiation resulted in a remarkable and constant increase in vitamin D2 content up to 60 minutes of exposure. Conversion at 254 nm was higher than at 312 nm reaching levels of 0.03 and 0.02 mg/g of vitamin D2, respectively, after 30 min of irradiation. Afterwards, longer irradiation times at 254 nm induced a slight decrease in vitamin D2 levels whereas at 312 nm did not further enhanced its biosynth

[ES]: El estilo de vida es un concepto complejo que incluye los aspectos externos a nosotros mismos que pueden modular e influir en nuestra salud. Los conocimientos sobre la relación entre el estilo de vida y el riesgo cardiovascular están lejos de proporcionar los niveles de evidencia que se han conseguido con los ensayos clínicos con fármacos, debido a que los estudios son escasos, fundamentalmente de tipo observacional y en grandes cohortes, con la dificultad añadida de la no existencia de métodos seguros para conocer con precisión la ingesta diaria o lo largo de tiempo, así como la dificultad en la recogida de datos y en la medición de la adherencia debido a diferencias en la composición de los alimentos en distintas épocas y a la conducta alimentaria cambiante que posee el ser humano a lo largo del tiempo. En este documento nos hemos propuesto llevar a cabo una revisión actualizada y jerarquizada en base a las evidencias actuales, prestando atención a tres aspectos que tienen gran importancia patogénica y que podemos modificar directamente: la actividad física, el consumo de tabaco y el patrón de alimentación. Con ello pretendemos actualizar los nuevos conocimientos sobre su relación con el riesgo cardiovascular aportando evidencias útiles, constituyendo una herramienta sencilla dirigida especialmente a todos los profesionales de la salud implicados en el cuidado de personas con riesgo cardiovascular para trasladarlos a la práctica clínica, definiendo unas líneas de actuación sencillas y fáciles para ser transmitidas a las personas que reciban un consejo para la prevención primaria y secundaria de las enfermedades cardiovasculares., [EN]: Lifestyle is a complex concept that includes aspects external to ourselves that can modulate and influence our health. The knowledge of the relationship between lifestyle and cardiovascular risk does not attain the level of evidence achieved with clinical trials with drugs, because clinical studies are scarce and mainly of observational nature, albeit based on large cohorts. Nutritional epidemiology has the added difficulty of being based mostly on subjective dietary recall methods to ascertain nutrient and food intake over time, with the additional problems of incomplete data collection, variable measurements of adherence due to seasonal and geographical differences in food composition, and the changing eating behavior that human beings have over time. The purpose of this document is to carry out an updated and hierarchical review of the relationship between lifestyle and cardiovascular disease based on current evidence, paying attention to three aspects that are of great pathogenic importance and are directly modifiable: physical activity, tobacco consumption, and diet. With this, we intend to update the knowledge on this relationship, construct evidence-based recommendations, and provide a simple tool for clinical practice especially directed to health professionals involved in the care of people at cardiovascular risk, defining simple and easy strategies for individuals who receive advice for the primary and secondary prevention of cardiovascular diseases.

The combination of supercritical fluid extraction followed by UV-irradiation resulted in an interesting strategy to obtain ergosterol- and Vitamin D2-enriched extracts from Lentinula edodes to design novel functional food. Extractions carried out following a specific experimental design pointed out that extraction yields and ergosterol concentrations were more influenced by pressure than by the extraction temperature although, high temperatures (55–75 °C) might induce transformation of ergosta-7,22-dienol and fungisterol into ergosterol. After extraction, the ergosterol-enriched extracts should be dissolved in methanol or ethanol and irradiated (25 °C, 4 cm) to partially transform ergosterol into vitamin D2. Irradiation at 365 nm was less effective than at 254 nm. The fastest transformation was obtained using a UV lamp covering the complete UV spectrum for a maximum of 1 h. However, this lamp also induced vitamin D4 formation although in lower amounts than vitamin D2 or lumisterol2 while with irradiation at 254 nm most of ergosterol was transformed into vitamin D2.

A literature and database review and several experiments were carried out because of the perturbing increase and exponential spreading of a major mistake thought scientific publications. Up to October 2015, more than 136 reports ensured that mushrooms contain flavonoids. However, 91% of the publications used an unspecific colorimetric method that was developed to quantify them in plants. However, no sequences encoding chalcone synthase or chalcone isomerase (key enzymes involved in the flavonoid biosynthetic pathway) were found in genomic databases even in completely sequenced mushrooms indicating that they do not have the enzymes needed for their production. Fruiting bodies obtained from substrates containing onion wastes and mycelia grown on media supplemented with specific flavonoids showed no flavonoid content after HPLC DAD/MS determinations indicating that they were also unable of absorbing them.

Culinary-medicinal mushrooms are able to lower blood cholesterol levels in animal models by different mechanisms. They might impair the endogenous cholesterol synthesis and exogenous cholesterol absorption during digestion. Mushroom extracts, obtained using pressurized water extractions (PWE) from Agaricus bisporus basidiomes, supplemented or not supplemented with selenium, were applied to HepG2 cell cultures to study the expression of 19 genes related to cholesterol homeostasis by low-density arrays (LDA). Only the PWE fractions obtained at 25°C showed 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) inhibitory activity. Besides the enzymatic inhibition, PWE extracts may downregulate some of the key genes involved in the cholesterol homeostasis, such as the squalene synthase gene (FDFT1), since its mRNA expression falls by one third of its initial value. In summary, A. bisporus extracts may also modulate biological cholesterol levels by molecular mechanisms further than the enzymatic way previously reported.

La presente invención se refiere a una bebida funcional que comprende vino desalcoholizado, aroma de vino, extracto de lúpulo enriquecido en isoxantohumol y, opcionalmente extracto de orujo de uva y extracto de semillas de uva, y a un procedimiento para la obtención del extracto de lúpulo, así como al uso de dicha bebida en la prevención o tratamiento del síndrome metabólico.

La presente invención se refiere a una bebida funcional que comprende vino desalcoholizado, aroma de vino, extracto de lúpulo enriquecido en isoxantohumol y, opcionalmente extracto de orujo de uva y extracto de semillas de uva, y a un procedimiento para la obtención del extracto de lúpulo, así como al uso de dicha bebida en la prevención o tratamiento del síndrome metabólico.

The methanol/water and particularly the water extracts obtained from 26 mushroom species were able to inhibit the 3-hydroxy-3-methyl-glutaryl CoA reductase (HMGCR) activity to different extent (10-76%). Cultivated mushrooms such as Pleurotus sp. and Lentinula edodes were among the strains which showed higher HMGCR inhibitory capacities. Their inhibitory properties were not largely influenced by cultivation parameters, mushroom developmental stage or flush number. The HMGCR inhibitory activity of L. edodes was concentrated in the cap excluding the gills while in Pleurotus ostreatus it was distributed through all the different tissues. A method to obtain aqueous fractions with high HMGCR inhibitory activity was optimized using an accelerated solvent extractor (ASE) by selecting 10.7. MPa and 25 °C as common extraction conditions and 5 cycles of 5. min each for P. ostreatus fruiting bodies and 15 cycles of 5. min for L. edodes suggesting that the potential HMGCR inhibitors are different in the two selected mushrooms. © 2012 Elsevier Ltd.

La presente invención se refiere a un procedimiento para la obtención de extracto de lúpulo enriquecido en isoxantohumol, y al extracto obtenido mediante dicho procedimiento.

One of the strategies in prevention or treatment of obesity is altering metabolism of lipids by inhibition of dietary fat absorption. The extracts obtained with methanol, water and methanol:water (1:1) from 21 mushroom species were screened as potential sources of pancreatic lipase (PL) inhibitors using a standardised in vitro test. Lepiota procera methanol:water (1:1) extracts showed the highest inhibition activity closely followed by Grifola frondosa, Pleurotus eryngii and Lyophyllum shimeji. Other mushroom strains such as Morchella conica, Marasmius oreades, Lentinula edodes, Amanita ponderosa and Boletus edulis also showed a certain inhibitory activity. However, when the PL inhibitory activity was evaluated using an in vitro digestion model mimicking gut conditions, none of the selected mushroom extracts were able to inhibit PL activity. On the contrary, stimulation of the lipase activity levels was observed and it was not due to endogenous mushroom lipases activities. © 2012 The Authors. International Journal of Food Science and Technology © 2012 Institute of Food Science and Technology.

[EN] The invention relates to a functional drink containing dealcoholised wine, aroma of wine, isoxanthohumol-enriched hops extract and, optionally, grape marc extract and grape seed extract. The invention also relates to a method for obtaining the hops extract and to the use of said drink in the prevention or treatment of metabolic syndrome., [ES] La presente invención se refiere a una bebida funcional que comprende vino desalcoholizado, aroma de vino, extracto de lúpulo enriquecido en isoxantohumol y, opcionalmente extracto de orujo de uva y extracto de semillas de uva, y a un procedimiento para la obtención del extracto de lúpulo, así como al uso de dicha bebida en la prevención o tratamiento del sindrome metabólico.

The anti-oxidant properties of liver pâtés might be enhanced by adding specific bioactive ingredients such as Boletus edulis mushrooms. Water- and methanol-soluble fractions of supplemented pâtés showed higher 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid and 2,2-diphenyl-1-picrylhydrazyl scavenging activities than control samples. B. edulis supplementation resulted in lower EC50 but only up to 5% (w/w). The high anti-oxidant activity observed in supplemented pâtés was stable during 30 storage days and decreased after 60 days but still it was higher than control pâtés and did not influence lipid oxidation. Addition of dried water or methanol preparations extracted from the mushroom did not improve the anti-oxidant activity observed when the complete fruiting body was utilized. Ergosterol-related compounds and ergotioneine were involved, but they were not exclusively responsible of the high anti-oxidant activity observed, phenolic compounds might be involved too.

Non-synonymous single nucleotide polymorphisms (nsSNPs) are genetic variations that affect the encoded protein by an amino acid change. In humans, many naturally-occurring nsSNPs cause protein dysfunction and increase vulnerability to disease. Identification of such nsSNPs provides an important opportunity to develop drugs/nutrients with precise therapeutic targets. Therefore, current biomedical research and medicinal chemistry look for targets and functional nsSNPs, to establish correlation with disease susceptibility and foster rational drug design. We review the molecular bases of missense mutation effects at the protein level, namely on sequence conservation, including stability, conformation, biophysical parameters, and protein-protein interaction. Further, we summarize some computational methods, available information resources, and the current approaches used to predict nsSNPs functionality in human genome, most of which based on protein structures and/or evolutionary conservation. Finally, using an approach paradigmatic of the nsSNPs-gene interactions, we evaluate the functional consequences and phenotypic effects of nsSNPs on two genes associated with cholesterol response. Biophysical changes produced by exchanged amino acids I638V (rs5908) from the 3-hydroxy-3-methylglutarylcoenzyme A reductase gene, and A370T (rs11669576) from the low density lipoprotein receptor gene have been analyzed with an emphasis on stability, activity, and structure of their related proteins. Based on available data and the results of our study, we propose that, even though the extent and precise nature of nsSNPs' role in health and disease is yet to be fully elucidated, targeted investigations are warranted and will - in the future - provide useful tools to develop targeted drugs. © 2011 Bentham Science Publishers.

Ligninolytic enzymes activities (laccases, peroxidases (total, MnP and MiP) and aryl-alcohol oxidase (AAO)) were measured during the cultivation of six commercial Pleurotus sp. strains on MMP media, on cereal grains (spawn) and on straw substrates (the three commonly utilized cultivation steps to obtain fruiting bodies) supplemented with several concentrations of autoclaved (OMW) or gamma-irradiated (iOMW) olive mill waste. Results indicated that all the strains were able to grow on MMP media and spawn containing up to 30% OMW and iOMW and on straw substrates mixed with 50% OMW. None of the strains showed AAO activity and there was not a single strain which showed the highest laccases and peroxidases activities, independently of the utilized substrate. Pleurotus mycelia adjusted their enzymatic mechanisms depending on their variety, type of substrate, concentration of OMW or iOMW added. OMW was a better supplement to use than iOMW because OMW induced higher exo-enzymes activities.

Seven Oyster mushroom strains were cultivated in wheat straw (WS) bags supplemented with 0 up to 90% olive mill waste (OMW), a solid residue obtained from a two-phases olive oil production system. All mushroom strains could grow but high OMW concentrations resulted in a significant yield, biological efficiency and productivity decrease, retarding of pinning and flushing and loss of fruiting bodies quality. However, most of the mushroom strains showed no significant differences on cultivation parameters and fruiting bodies quality (except for colour) between control (WS) substrates or substrates supplemented up to 50% OMW (w/w). Addition of 50% OMW to WS substrates turned mushrooms lighter and less reddish. This colour shift was not due to enzymatic oxidations or inhibition since laccase and peroxidase activities were not differing from control. Total phenolic content and antioxidant activity were also similar and no phenolic compounds from OMW were detected in the fruiting bodies. Some of the analysed Pleurotus strains were better suited to grow on OMW supplemented substrates than others.

A new procedure has been developed to separate and characterize antioxidant compounds from Spirulina platensis microalga based on the combination of pressurized liquid extraction (PLE) and different chromatographic procedures, such as Thin-Layer Chromatography (TLC), at preparative scale, and High Performance Liquid Chromatography (HPLC) with a Diode Array Detector (DAD). Different solvents were tested for PLE extraction of antioxidants from Spirulina platensis microalga. An optimized pressurized liquid extraction process using ethanol (generally recognized as safe, GRAS) as extraction solvent has been obtained that provides natural extracts with high yields and good antioxidant properties. TLC analysis of this ethanolic extract obtained at 115°C for 15 min was carried out and the silica layer was stained with a DPPH• (diphenyl-pycril-hydrazyl) radical solution to determine the antioxidant activity of the different chromatographic bands. Next, these coloured bands were collected for their subsequent analysis by HPLC-DAD revealing that the compounds with the most important antioxidant activity present in Spirulina extracts were carotenoids, as well as phenolic compounds and degradation products of chlorophylls.

[EN]Reaction occurs in a medium buffered with phosphate in an aqueous medium, at neutral pH and ambient temperature. Hence, the reaction medium consists of: tyrosol as precursor; (commercial) mushroom tyrosinase to catalyze the process and vitamin C in excess. Reaction starts after initial shaking. The reaction stops when the concentration of the initial tyrosol is exhausted. In order for reaction to continue, it will suffice to add more tyrosol (always ensuring that the vitamin C/tyrosol ratio is more than 1). Once the desired concentration of hydroxytyrosol, it is filtered. The enzyme with molecular weight over twice the size of the pore is retained in the filter and can be used once again. This first extract, which is very enriched in antioxidant hydroxytyrosol with a high antioxidant capacity (in the present case, in combination with vitamin C), can already be used as food additive. [ES]La reacción transcurre en un medio tamponado con fosfato en medio acuoso, a pH neutro, y a temperatura ambiente. El medio de reacción, se compone por tanto de: tirosol, como precursor tirosinasa de champiñón (comercial) para catalizar el proceso y vitamina C en exceso. Tras una agitación inicial, la reacción comienza. Cuando la concentración de tirosol inicial se agote la reacción se para. Para que continúe basta con añadir más tirosol (siempre cuidando que la relación vitamina C/ tirosol sea mayor que 1). Cuando se obtenga la concentración de hidroxitirosol deseada, se filtra. La enzima, de peso molecular más del doble del tamaño de poro, queda retenida en el filtro, y puede ser de nuevo reutilizada. Este primer extracto muy enriquecido en antioxidante hidroxitirosol, con una alta capacidad antioxidante, (en este caso, en combinación con vitamina C), ya es susceptible de utilizarse como aditivo alimentario.

Hydroxytyrosol (HTyr), a natural ortho-diphenolic antioxidant with health-beneficial properties that mainly occurs in virgin olive oil and olive oil mill waste waters (also known as vegetative waters), has been enzymatically synthesized using mushroom tyrosinase. This o-diphenol (not commercially available) was obtained from its monophenolic precursor tyrosol (commercially available) in the presence of both tyrosinase and ascorbic acid. The reaction synthesis is continuous, easy to perform, and adaptable to a bioreactor for industrial purposes. The HTyr concentration is time-predicted, and the yield of reaction can be 100%. The synthesis method reported here is an alternative approach to obtain this compound in an environmentally friendly way.

An easy and fast test has been designed to compare the total free radical scavenging capacity (RSC) of various food samples. Black and green teas from different countries, and wines of different brands were studied and compared as examples of coloured liquids (water- and methanol-soluble); oils from different sources were used as examples of lipidic foodstuffs; apples of different varieties and spinach were analysed as solid foods. Dilutions of extracts of the described foodstuffs were prepared and aliquots of each dilution were spotted onto TLC silica gel layers in the form of a dot-blot test: layers were stained with a methanolic solution of the 2,2-diphenyl-1-picrylhydrazyl radical. Dots of extracts of foodstuffs with RSC turned yellow, with a colour intensity depending on the RSC compounds present in the dilutions. After 1 h staining, the intensity of the yellow colour was measured with a chromameter (b* parameter) at one of the dilutions at which the colour value was linearly correlated to the concentration of the sample. According to these readings the different samples were organised in decreasing order of b* values, an order which corresponded to the decreasing order of RSC as determined by spectrophotometric methods. The dot-blot test was sensitive enough to detect differences of RSC between varieties and brands of water- or methanol-soluble products, but was not adequate for lipid-based compounds. The test was also able to follow the variation of RSC during food processing as in, for example, the heat-treatment of spinach.

The total free radical scavenger capacity (RSC) of 57 edible oils from different sources was studied: olive (24 brands of oils), sunflower (6), safflower (2), rapeseed (3), soybean (3), linseed (2), corn (3), hazelnut (2), walnut (2), sesame (2), almond (2), mixture of oils for salad (2), “dietetic” oil (2), and peanut (2). Olive oils were also studied according to their geographical origins (France, Greece, Italy, Morocco, Spain, and Turkey). RSC was determined spectrophotometrically by measuring the disappearance of the radical 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•) at 515 nm. The disappearance of the radical followed a double-exponential equation in the presence of oils and oil fractions, which suggested the presence of two (fast and slow) groups of antioxidants. RSC was studied for the methanol-soluble phase (“methanolic fraction”, MF) of the oil, the fraction nonsoluble in methanol (“lipidic fraction”, LF), and the nonfractionated oil (“total oil”; TF = MF + LF). Only olive, linseed, rapeseed, safflower, sesame, and walnut oils showed significant RSC in the MF due to the presence of phenolic compounds. No significant differences were found in the RSC of olive oils from different geographical origins. Upon heating at 180 °C the apparent constant for the disappearance of RSC (kT) and the half-life (t1/2) of RSC for MF, LF, and TF were calculated. The second-order rate constants (k2) for the antiradical activity of some phenolic compounds present in oils are also reported.

In this paper, oleuropein and some other related phenolic compounds are reviewed. Their occurrence, distribution, biosynthesis and transformation during maturation and during industrial processing (preparation of table olives and oil production) are described. Their role in human health is proposed based on current human, animal and in vitro studies as molecules with antioxidant and antimicrobial properties.