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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.