Exploring the potential of phenolic compounds from the coffee pulp in preventing cellular oxidative stress after in vitro digestion.

The coffee pulp, a by-product of the coffee industry, contains a high concentration of phenolic compounds and caffeine. Simulated gastrointestinal digestion may influence these active compounds' bioaccessibility, bioavailability, and bioactivity. Understanding the impact of the digestive metabolism on the coffee pulp's phenolic composition and its effect on cellular oxidative stress biomarkers is essential. In this study, we evaluated the influence of in vitro gastrointestinal digestion of the coffee pulp flour (CPF) and extract (CPE) on their phenolic profile, radical scavenging capacity, cellular antioxidant activity, and cytoprotective properties in intestinal epithelial (IEC-6) and hepatic (HepG2) cells. The CPF and the CPE contained a high amount of caffeine and phenolic compounds, predominantly phenolic acids (3',4'-dihydroxycinnamoylquinic and 3,4-dihydroxybenzoic acids) and flavonoids (3,3',4',5,7-pentahydroxyflavone derivatives). Simulated digestion resulted in increased antioxidant capacity, and both the CPF and the CPE demonstrated free radical scavenging abilities even after in vitro digestion. The CPF and the CPE did not induce cytotoxicity in intestinal and hepatic cells, and both matrices exhibited the ability to scavenge intracellular reactive oxygen species. The coffee pulp treatments prevented the decrease of glutathione, thiol groups, and superoxide dismutase and catalase enzymatic activities evoked by tert-butyl hydroperoxide elicitation in IEC-6 and HepG2 cells. Our findings suggest that the coffee pulp could be used as a potent food ingredient for preventing cellular oxidative stress due to its high content of antioxidant compounds.

Radical Scavenging and Cellular Antioxidant Activity of the Cocoa Shell Phenolic Compounds after Simulated Digestion.

The cocoa industry generates a considerable quantity of cocoa shell, a by-product with high levels of methylxanthines and phenolic compounds. Nevertheless, the digestion process can extensively modify these compounds' bioaccessibility, bioavailability, and bioactivity as a consequence of their transformation. Hence, this work's objective was to assess the influence of simulated gastrointestinal digestion on the concentration of phenolic compounds found in the cocoa shell flour (CSF) and the cocoa shell extract (CSE), as well as to investigate their radical scavenging capacity and antioxidant activity in both intestinal epithelial (IEC-6) and hepatic (HepG2) cells. The CSF and the CSE exhibited a high amount of methylxanthines (theobromine and caffeine) and phenolic compounds, mainly gallic acid and (+)-catechin, which persisted through the course of the simulated digestion. Gastrointestinal digestion increased the antioxidant capacity of the CSF and the CSE, which also displayed free radical scavenging capacity during the simulated digestion. Neither the CSF nor the CSE exhibited cytotoxicity in intestinal epithelial (IEC-6) or hepatic (HepG2) cells. Moreover, they effectively counteracted oxidative stress triggered by tert-butyl hydroperoxide (t-BHP) while preventing the decline of glutathione, thiol groups, superoxide dismutase, and catalase activities in both cell lines. Our study suggests that the cocoa shell may serve as a functional food ingredient for promoting health, owing to its rich concentration of antioxidant compounds that could support combating the cellular oxidative stress associated with chronic disease development.

Changes in the cocoa shell dietary fiber and phenolic compounds after extrusion determine its functional and physiological properties.

The influence of different extrusion conditions on the cocoa shell (CS) dietary fiber, phenolic compounds, and antioxidant and functional properties was evaluated. Extrusion produced losses in the CS dietary fiber (3-26%), especially in the insoluble fraction, being more accentuated at higher temperatures (160 °C) and lower moisture feed (15-20%). The soluble fiber fraction significantly increased at 135 °C because of the solubilization of galactose- and glucose-containing insoluble polysaccharides. The extruded CS treated at 160 °C-25% of feed moisture showed the highest increase of total (27%) and free (58%) phenolic compounds, accompanied by an increase of indirect (10%) and direct (77%) antioxidant capacity. However, more promising results relative to the phenolic compounds' bioaccessibility after in vitro simulated digestion were observed for 135°C-15% of feed moisture extrusion conditions. The CS' physicochemical and techno-functional properties were affected by extrusion, producing extrudates with higher bulk density, a diminished capacity to hold oil (22-28%) and water (18-65%), and improved swelling properties (14-35%). The extruded CS exhibited increased glucose adsorption capacity (up to 2.1-fold, at 135 °C-15% of feed moisture) and α-amylase in vitro inhibitory capacity (29-54%), accompanied by an increase in their glucose diffusion delaying ability (73-91%) and their starch digestion retardation capacity (up to 2.8-fold, at 135 °C-15% of feed moisture). Moreover, the extruded CS preserved its cholesterol and bile salts binding capacity and pancreatic lipase inhibitory properties. These findings generated knowledge of the CS valorization through extrusion to produce foods rich in dietary fiber with improved health-promoting properties due to the extrusion-triggered fiber solubilization.

Valorization of coffee pulp as bioactive food ingredient by sustainable extraction methodologies.

Coffee pulp is an underutilized by-product of coffee industrial production rich in bioactive compounds such as phenolic compounds, caffeine, and dietary fiber. The widely known antioxidant, anti-inflammatory, anti-aging, antimicrobial and hepatoprotective health-promoting properties attributed to mentioned compounds enhance the use of coffee pulp as a bioactive food ingredient. Furthermore, the application of green sustainable extraction techniques pursuing highly efficient and selective extraction processes promotes this by-product exploitation in food science. Hence, this review gathers the available information relative to the impact of the extraction processes on the bioactive compound's recovery from coffee pulp, providing an overview of the most recent advances. An in-depth comparison workout between conventional and alternative extraction methods was performed to identify the most suitable techniques for coffee pulp valorization as functional ingredient until date. A critical discussion focused on advantages and drawbacks of the extraction methods applied to coffee pulp was included together a prospective of emerging extraction techniques.

Slower Growth during Lactation Rescues Early Cardiovascular and Adipose Tissue Hypertrophy Induced by Fetal Undernutrition in Rats.

Low birth weight (LBW) and accelerated growth during lactation are associated with cardiometabolic disease development. LBW offspring from rats exposed to undernutrition during gestation (MUN) develops hypertension. In this rat model, we tested if slower postnatal growth improves early cardiometabolic alterations. MUN dams were fed ad libitum during gestation days 1-10, with 50% of the daily intake during days 11-21 and ad libitum during lactation. Control dams were always fed ad libitum. Pups were maintained with their own mother or cross-fostered. Body weight and length were recorded weekly, and breastmilk was obtained. At weaning, the heart was evaluated by echocardiography, and aorta structure and adipocytes in white perivascular fat were studied by confocal microscopy (size, % beige-adipocytes by Mitotracker staining). Breastmilk protein and fat content were not significantly different between groups. Compared to controls, MUN males significantly accelerated body weight gain during the exclusive lactation period (days 1-14) while females accelerated during the last week; length growth was slower in MUN rats from both sexes. By weaning, MUN males, but not females, showed reduced diastolic function and hypertrophy in the heart, aorta, and adipocytes; the percentage of beige-type adipocytes was smaller in MUN males and females. Fostering MUN offspring on control dams significantly reduced weight gain rate, cardiovascular, and fat hypertrophy, increasing beige-adipocyte proportion. Control offspring nursed by MUN mothers reduced body growth gain, without cardiovascular modifications. In conclusion, slower growth during lactation can rescue early cardiovascular alterations induced by fetal undernutrition. Exclusive lactation was a key period, despite no modifications in breastmilk macronutrients, suggesting the role of bioactive components. Our data support that lactation is a key period to counteract cardiometabolic disease programming in LBW and a potential intervention window for the mother.

Influence of Neonatal Sex on Breast Milk Protein and Antioxidant Content in Spanish Women in the First Month of Lactation.

Breast milk (BM) is the best food for newborns. Male sex is associated with a higher risk of fetal programming, prematurity, and adverse postnatal outcome, being that BM is an important health determinant. BM composition is dynamic and modified by several factors, including lactation period, prematurity, maternal nutritional status, and others. This study was designed to evaluate the influence of sex on BM composition during the first month of lactation, focused on macronutrients and antioxidants. Forty-eight breastfeeding women and their fifty-five newborns were recruited at the Hospital Clínico San Carlos (Madrid, Spain). Clinical sociodemographic data and anthropometric parameters were collected. BM samples were obtained at days 7, 14, and 28 of lactation to assess fat (Mojonnier method), protein (Bradford method), and biomarkers of oxidative status: total antioxidant capacity (ABTS and FRAP methods), thiol groups, reduced glutathione, superoxide dismutase and catalase activities, lipid peroxidation, and protein oxidation (spectrophotometric methods). Linear mixed models with random effects adjusted by maternal anthropometry, neonatal Z-scores at birth, and gestational age were used to assess the main effects of sex, lactation period, and their interaction. BM from mothers with male neonates exhibited significantly higher protein, ABTS, FRAP, and GSH levels, while catalase showed the opposite trend. No differences between sexes were observed in SOD, total thiols, and oxidative damage biomarkers. Most changes were observed on day 7 of lactation. Adjusted models demonstrated a significant association between male sex and proteins (ß = 2.70 ± 1.20; p-Value = 0.048). In addition, total antioxidant capacity by ABTS (ß = 0.11 ± 0.06) and GSH (ß = 1.82 ± 0.94) showed a positive trend near significance (p-Value = 0.056; p-Value = 0.064, respectively). In conclusion, transitional milk showed sex differences in composition with higher protein and GSH levels in males. This may represent an advantage in the immediate perinatal period, which may help to counteract the worse adaptation of males to adverse intrauterine environments and prematurity.

Pressurized carbon dioxide as a potential tool for decellularization of pulmonary arteries for transplant purposes.

Vascular bio-scaffolds produced from decellularized tissue offer a promising material for treatment of several types of cardiovascular diseases. These materials have the potential to maintain the functional properties of the extracellular matrix (ECM), and allow for growth and remodeling in vivo. The most commonly used methods for decellularization are based on chemicals and enzymes combinations, which often damage the ECM and cause cytotoxic effects in vivo. Mild methods involving pressurized CO2-ethanol (EtOH)-based fluids, in a supercritical or near supercritical state, have been studied for decellularization of cardiovascular tissue, but results are controversial. Moreover, data are lacking on the amount and type of lipids remaining in the tissue. Here we show that pressurized CO2-EtOH-H2O fluids (average molar composition, ΧCO2 0.91) yielded close to complete removal of lipids from porcine pulmonary arteries, including a notably decrease of pro-inflammatory fatty acids. Pressurized CO2-limonene fluids (ΧCO2 0.88) and neat supercritical CO2 (scCO2) achieved the removal of 90% of triacylglycerides. Moreover, treatment of tissue with pressurized CO2-limonene followed by enzyme treatment, resulted in efficient DNA removal. The structure of elastic fibers was preserved after pressurized treatment, regardless solvent composition. In conclusion, pressurized CO2-ethanol fluids offer an efficient tool for delipidation in bio-scaffold production, while pressurized CO2-limonene fluids facilitate subsequent enzymatic removal of DNA.

Strengths and weaknesses of the aniline-blue method used to test mushroom (1→3)-ß-d-glucans obtained by microwave-assisted extractions.

The parameters to extract polysaccharide-enriched fractions (PEF) from mushrooms using MAE (microwave-assisted extraction) were adjusted following a full factorial 32 experimental design. The highest yield and total carbohydrate values, using Lentinula edodes as model mushroom, were obtained at 180 °C and 30 min. Several mushroom species were submitted to MAE and their PEF yields ranged between 12.1-44.2%. (1→3)-ß-Glucans determination using a conventional fluorimetric method changed depending on the standard utilized. NMR analyses of PEF indicated that the presence of other polysaccharides in the extracts or their specific folding, might impair the proper determination of (1→3) linkages by the fluorophore. Mushrooms from Cantharellales order contained (1→3)-ß-glucans but they were not detected with the fluorimetric method. Therefore, although the method (after adjustments) was sensitive enough to detect their presence in many mushroom extracts, it cannot be used for all species and it is also not recommended for quantitative determinations.

Efficient methodology for the extraction and analysis of lipids from porcine pulmonary artery by supercritical fluid chromatography coupled to mass spectrometry.

Pulmonary artery grafts are needed as cardiovascular bioprosthetics. For successful tissue recellularization after transplantation, lipids have to be removed from the donor artery. Developing a selective process to remove lipids without damaging the extracellular matrix greatly depends on knowing the amount and type of lipid compounds in the specific tissue. Here we present an efficient methodology for the study of lipids present in porcine pulmonary arteries. The performance of six extraction methods to recover lipids from artery was evaluated. For this purpose, a supercritical fluid chromatography method coupled to quadrupole time-of-flight mass spectrometry detection (UHPSFC/QTOF-MS) was adapted. The method enabled separation of lipids of a wide range of polarity according to lipid class in less than 7 min. One dichloromethane-based extraction method was shown to be the most efficient one for the recovery of lipids from pulmonary artery. However, one MTBE-based extraction method was able to show the highest fatty acid extraction yields (to the expense of longer extraction times). Lipids were relative quantified according to class, and the major species within each class were identified. Triacylglycerols and glycerophospholipids were the most abundant classes, followed by sphingomyelins, monoacylglycerols and fatty acyls. The matrix effect exerted no interference on the analytical method, except for some few combinations of extraction method and lipid class. These results are of relevance for lipidomic studies from solid tissue, in particular for studies on pulmonary and cardiovascular diseases. Finally, our work sets the basis for the further development of a selective processes to remove lipids from pulmonary artery without damaging the tissue prior to transplantation.

Data on saponins, xylan and cellulose yield obtained from quinoa stalks after pressurized hot water extraction.

The data we present below are linked to our research paper "Integrated process for sequential extraction of saponins, xylan and cellulose from quinoa stalks (Chenopodium quinoa Willd.)" (Gil-Ramírez et al., 2018) [1]. The objective is to provide supplementary information in order to facilitate the comprehension of the central composite experimental design (rotatable 22) used in the integrated process of extractions. Two factors, temperature and time of extraction are considered in the design. The responses are the yield of saponin, xylan and cellulose. First, the desirable linear regression obtained by the observed vs. predicted yields plot for each variable response confirm the validation of the model (Fig. 1). Second, the data presented here through Standardized Pareto Charts (Fig. 2), provides information about the effect of the time and temperature, as well as their interactions, in the yield of saponins, xylan and cellulose obtained in an integrated sequential extraction.

Molecular actions of hypocholesterolaemic compounds from edible mushrooms.

Cholesterol levels are strictly regulated to maintain its homeostasis; therefore, if it is not absorbed with the diet, the cholesterol biosynthetic pathway is enhanced and vice versa. Nowadays, the commonly prescribed therapeutic treatments for hypocholesterolemic patients are targeted toward the reduction of both cholesterol intestinal absorption and/or its endogenous biosynthesis. But, when hypercholesterolemia is still moderate the consumption of food products with cholesterol-lowering capacities is more desirable than using drugs. Marketed foods supplemented with hypocholesterolemic compounds are only inhibiting mechanisms for cholesterol absorption (i.e. phytosterols and cereal ß-glucans). However, certain fungal extracts obtained from edible mushrooms might be able to modulate cholesterol levels by both strategies, pharmaceutical drugs and functional foods. In vitro and in vivo studies indicated that fungal sterols down-regulated genes involved in cholesterol homeostasis (such as Srebf2 and Nr1h4 (FXR)) and other specific mushroom extracts (ß-glucans and other water-soluble compounds) also stimulated transcriptional profiles similar to simvastatin or ezetimibe (two hypocholesterolemic drugs). These and other observations suggested that the hypocholesterolemic effect of mushroom extracts could be due to transcriptional and post-transcriptional modulations besides other indirect effects.

Water-Soluble Polysaccharide Extracts from the Oyster Culinary-Medicinal Mushroom Pleurotus ostreatus (Agaricomycetes) with HMGCR Inhibitory Activity.

Water extracts from Pleurotus ostreatus containing no statins showed 3-hydroxy-3-methyl-glutaryl CoA reductase (HMGCR) inhibitory activity (in vitro) that might be due to specific water-soluble polysaccharides (WSPs); when isolated and deproteinized, increasing concentrations of the WSP extract induced higher inhibition. The WSP extract contained mainly ß-glucans, mannogalactans, and glycogen (e.g., α-glucans), although derivatives or fragments with lower molecular weights (between 14 and 3.5 kDa) were present and were able to induce the inhibitory activity. The extract contained more ß-(1→3)-glucans than ß-(11→3),(11→6)-glucans, and they partially survived digestion and managed to pass through Caco2 cell monolayers to the lower compartment after in vitro digestion and transport experiments. The WSP might also modulate Caco2 membrane integrity.

Evaluation of microwave-assisted and pressurized liquid extractions to obtain ß-d-glucans from mushrooms.

Microwave-assisted extraction (MAE) and pressurized liquid extraction (PLE) were compared as advanced technologies to obtain polysaccharides (particularly biologically active ß-glucans) from Pleurotus ostreatus and Ganoderma lucidum fruiting bodies. Extraction effectiveness was compared by a full-factorial experimental design (response surface methodology, RSM), using water as extraction solvent. Total carbohydrate content of the obtained extracts and polysaccharide yields were the variable responses investigated, while temperature and extraction time were the experimental factors. Temperature showed stronger influence in the polysaccharide extraction than time. The latter factor slightly affected MAE but not PLE extractions. Optimal conditions within the studied range were determined for each extraction method and species based on the desirability functions. Regarding the polysaccharide composition, the main differences between the species were more quantitative rather than qualitative, since NMR analyses indicated that all extracts contained mainly ß- and α-glucans and heteropolysaccharides. Both extraction systems were effective for polysaccharide extraction from mushrooms.

Water-Soluble Compounds from Lentinula edodes Influencing the HMG-CoA Reductase Activity and the Expression of Genes Involved in the Cholesterol Metabolism.

A water extract from Lentinula edodes (LWE) showed HMG-CoA reductase inhibitory activity but contained no statins. NMR indicated the presence of water-soluble α- and ß-glucans and fucomannogalactans. Fractions containing derivatives of these polysaccharides with molecular weight down to approximately 1 kDa still retained their inhibitory activity. Once digested LWE was applied to Caco2 in transport experiments, no significant effect was noticed on the modulation of cholesterol-related gene expression. But, when the lower compartment of the Caco2 monolayer was applied to HepG2, some genes were modulated (after 24 h). LWE was also administrated to normo- and hypercholesterolemic mice, and no significant lowering of serum cholesterol levels was observed; but reduction of triglycerides in liver was observed. However, LWE supplementation modulated the transcriptional profile of some genes involved in the cholesterol metabolism similarly to simvastatin, suggesting that it could hold potential as a hypolipidemic/hypocholesterolemic extract, although further dose-dependent studies should be carried out.

Plasma Cholesterol-Lowering Activity of Lard Functionalized with Mushroom Extracts Is Independent of Niemann-Pick C1-like 1 Protein and ABC Sterol Transporter Gene Expression in Hypercholesterolemic Mice.

Interest in food matrices supplemented with mushrooms as hypocholesterolemic functional foods is increasing. This study was to (i) investigate the hypocholesterolemic activity of lard functionalized with mushroom extracts (LF) including fungal ß-glucans, water-soluble polysaccharides, or ergosterol and (ii) examine the LF influence on transcriptional mechanisms involved in cholesterol metabolism. mRNA levels of 17 cholesterol-related genes were evaluated in jejunum, cecum, and liver of high cholesterol-fed mice. The four tested LFs decreased plasma cholesterol by 22-42%, HDLc by 18-40%, and LDLc by 27-51%, and two of them increased mRNA levels of jejunal Npc1l1 and Abcg5 and hepatic Npc1l1. mRNA levels of other cholesterol-related genes were unchanged. These findings suggest that LF may have potential as a dietary supplement for counteracting diet-induced hypercholesterolemia and could be a source for the development of novel cholesterol-lowering functional foods. However, the cholesterol-lowering effect was unrelated to transcriptional changes, suggesting that post-transcriptional mechanisms could be involved.

Modulation of cholesterol-related gene expression by ergosterol and ergosterol-enriched extracts obtained from Agaricus bisporus.

PURPOSE: To investigate the effect of two extracts obtained from Agaricus bisporus on the mRNA expression of cholesterol-related genes. One of the extracts contained ergosterol and other fungal sterols (SFE) and the other contained ß-glucans and fungal sterols (EßG). METHODS: Firstly, the dietary mixed micelles (DMMs) generated after in vitro digestion of standards and SFE were applied to Caco2 cells. Then, the lower compartment after a Caco2-transport experiment was applied to HepG2 cells. The mRNA expression was assessed in both cell lines by low-density arrays (LDA). Mice received the extracts, ergosterol or control drugs after 4 weeks of a high-cholesterol diet. The lipid profile of plasma, liver and feces was determined. LDA assays were performed in liver and intestines. RESULTS: The DMM fraction of SFE up-regulated the LDLR mRNA expression in Caco2 cells. The lower compartment after Caco2-transport experiments up-regulated LDLR and modulated several other lipid-related genes in HepG2 cells. In mice, SFE decreased TC/HDL ratio and reduced hepatic triglycerides paralleled with down-regulation of Dgat1 expression, while EßG did it without transcriptional changes. Addition of SFE or ergosterol induced in jejunum a similar transcriptional response to simvastatin and ezetimibe; they all down-regulated Srebf2 and Nr1h4 (FXR) genes. CONCLUSION: Ergosterol-containing extracts from A. bisporus lowered hepatic triglyceride and modify the mRNA expression of cholesterol-related genes although the transcriptional regulation was unrelated to changes in plasma lipid profile. These extracts may be useful limiting hepatic steatosis and as bioactive ingredients to design novel functional foods preventing lifestyle-related diseases such as non-alcoholic fatty liver disease.

Modulation of Cholesterol-Related Gene Expression by Dietary Fiber Fractions from Edible Mushrooms.

Mushrooms are a source of dietary fiber (DF) with a cholesterol-lowering effect. However, their underlying mechanisms are poorly understood. The effect of DF-enriched fractions from three mushrooms species on cholesterol-related expression was studied in vitro. The Pleurotus ostreatus DF fraction (PDF) was used in mice models to assess its potential palliative or preventive effect against hypercholesterolemia. PDF induced a transcriptional response in Caco-2 cells, suggesting a possible cholesterol-lowering effect. In the palliative setting, PDF reduced hepatic triglyceride likely because Dgat1 was downregulated. However, cholesterol-related biochemical data showed no changes and no relation with the observed transcriptional modulation. In the preventive setting, PDF modulated cholesterol-related genes expression in a manner similar to that of simvastatin and ezetimibe in the liver, although no changes in plasma and liver biochemical data were induced. Therefore, PDF may be useful reducing hepatic triglyceride accumulation. Because it induced a molecular response similar to hypocholesterolemic drugs in liver, further dose-dependent studies should be carried out.

Effect of Selenium-Enriched Agaricus bisporus (Higher Basidiomycetes) Extracts, Obtained by Pressurized Water Extraction, on the Expression of Cholesterol Homeostasis Related Genes by Low-Density Array.

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.

Pressurized water extraction of ß-glucan enriched fractions with bile acids-binding capacities obtained from edible mushrooms.

A pressurized water extraction (PWE) method was developed in order to extract ß-glucans with bile acids-binding capacities from cultivated mushrooms (Agaricus bisporus, Lentinula edodes, and Pleurotus ostreatus) to be used as supplements to design novel foods with hypocholesterolemic properties. Extraction yields were higher in individual than sequential extractions being the optimal extraction parameters: 200°C, 5 cycles of 5 min each at 10.3 MPa. The crude polysaccharide (PSC) fractions, isolated from the PWE extracts contained mainly ß-glucans (including chitooligosaccharides deriving from chitin hydrolysis), α-glucans, and other PSCs (hetero-/proteo-glucans) depending on the extraction temperature and mushroom strain considered. The observed bile acids-binding capacities of some extracts were similar to a ß-glucan enriched fraction obtained from cereals.

Study on the 3-hydroxy-3-methyl-glutaryl CoA reductase inhibitory properties of Agaricus bisporus and extraction of bioactive fractions using pressurised solvent technologies.

BACKGROUND: Agaricus bisporus mushrooms were able to lower cholesterol levels in hypercholesterolaemic rats and it was suggested that dietary fibre might inhibit cholesterol absorption. However, A. bisporus extracts were also able to inhibit the 3-hydroxy-3-methyl-glutaryl CoA reductase (HMGCR, the key enzyme in the cholesterol biosynthetic pathway) and this might also contribute to the observed lowering of cholesterol levels in serum. RESULTS: The methanol-water extracts obtained from A. bisporus were able to inhibit up to 60% the HMGCR activity using an in vitro assay. The HMGCR inhibitory capacities depended on cultivation conditions, strains, etc. The potential inhibitors were not statins, they might be ß-glucans able to scavenge the substrate and impair the enzymatic reaction. They were present during all mushroom developmental stages and similarly distributed through all the tissues including the parts discarded as a by-product. Accelerated solvent extractions using 1:1 ethanol-water as pressurised solvent (10.7 MPa, 25°C, five cycles of 5 min) were more effective in the extraction of the HMGCiR inhibitor(s) than supercritical fluid extractions (9 MPa, 40°C) using CO2 with 10% ethanol. CONCLUSION: A mushroom cultivation and two extraction procedures were optimised to obtain fractions from A. bisporus with high HMGCR inhibitory activities to design novel ingredients for hypocholesterolaemic functional foodstuffs.