Concentrations Of Ergosterol Research Articles

A Multi-Year Study of Mycotoxin Co-Occurrence in Wheat and Corn Grown in Ontario, Canada.

Mycotoxin emergence and co-occurrence trends in Canadian grains are dynamic and evolving in response to changing weather patterns within each growing season. The mycotoxins deoxynivalenol and zearalenone are the dominant mycotoxins detected in grains grown in Eastern Canada. Two potential emerging mycotoxins of concern are sterigmatocystin, produced by Aspergillus versicolor, and diacetoxyscirpenol, a type A trichothecene produced by a number of Fusarium species. In response to a call from the 83rd Joint Expert Committee on Food Additives and Contaminants, we conducted a comprehensive survey of samples from cereal production areas in Ontario, Canada. Some 159 wheat and 160 corn samples were collected from farms over a three-year period. Samples were extracted and analyzed by LC-MS/MS for 33 mycotoxins and secondary metabolites. Ergosterol was analyzed as an estimate of the overall fungal biomass in the samples. In wheat, the ratio of DON to its glucoside, deoxynivalenol-3-glucoside (DON-3G), exhibited high variability, likely attributable to differences among cultivars. In corn, the ratio was more consistent across the samples. Sterigmatocystin was detected in some wheat that had higher concentrations of ergosterol. Diacetoxyscirpenol was not detected in either corn or wheat over the three years, demonstrating a low risk to Ontario grain. Overall, there was some change to the mycotoxin profiles over the three years for wheat and corn. Ongoing surveys are required to reassess trends and ensure the safety of the food value chain, especially for emerging mycotoxins.

Changes in the Characteristics of Pine Logging Residue during Storage in Forest Stands

A significant amount of logging residue is produced during roundwood harvesting. Logs are often left in forest sites due to, for example, ecological or logistical aspects. Taking into account the fact that the number of studies focused on changes in the properties of the residue is very low, it was decided to conduct research on the effect of a three-year storage period in forest stands on the chemical properties and energy potential of the wood. The research design allowed for the determination of changes during up to three years of storage. The performed analysis showed a highly negative impact on the characteristics of the material. These adverse effects were probably caused by the activity of microorganisms such as fungi and bacteria, as evidenced by the increased concentrations of ergosterol and low-molecular-weight organic acids. Moreover, it was found that wood stored for three years was characterized by a lower cellulose content; an increased percentage of lignin; a reduced content of sterols (desmosterol, stigmasterol, lanosterol and β-sitosterol) and phenolic acids; and reduced antioxidant activity, as determined with the ABTS+ method. Storing logs also led to a reduction in the energy potential of the residues, as shown by a reduction in net and gross calorific value and an increased ash content.

Unveiling the power of adding sterols in wine: Optimizing alcoholic fermentation with strategic management

Excessive grape must clarification can result in sluggish alcoholic fermentation and sometimes alcoholic stuck fermentation, because of the lack of sterols for the yeasts growth. To avoid this risk, addition of sterols (ergosterol or phytosterols) can be performed, resulting in higher Saccharomyces cerevisiae viability and shorter fermentation duration. However, no dedicated study was implemented to evaluate the efficiency of different strategies of sterol management (considering sterol type, added concentration and timing of addition) during wine fermentation. So, first, to evaluate whether the response of wine yeast strains to sterol nutrition was similar according to the type and the concentration of the sterol present in the initial must, it was studied the response of a set of 10 S. cerevisiae strains in a synthetic grape must with low, medium and high concentrations of ergosterol or phytosterols. Then, the impact of the timing of sterol addition was evaluated on 2 S. cerevisiae strains with opposite behaviours. This work confirmed previous results concerning the role of ergosterol and phytosterols but also revealed new findings in this field. At first, it was confirmed that ergosterol played an important role in improving the maintenance of viable cells towards the end of fermentation, particularly in sterols-limited situations, while phytosterols demonstrated an ability to reduce acetate and glycerol production. But, in a second part, our study sheds new light on the beneficial impact of sterol addition on amino acid assimilation in yeast, leading to an increase in maximum fermentation rate, biomass production and percentage of viable cells. However, the main novelty of this research work concerns the timing of ergosterol addition. This addition at the start of fermentation in a phytosterols-free synthetic must enabled faster fermentations, as well as higher fermentative aroma synthesis, compared to addition during stationary phase Even if the impact of ergosterol additions were relatively similar for both strains tested, notable differences were found concerning amino acid assimilation and biomass production, suggesting differences in the regulation of nitrogen metabolism between both strains. These findings provide new insights into our understanding of sterol role in enological fermentation. It offers a basis for both the development of innovative strategies for sterol management and the selection of wine yeast strains under sterol starvation.

Impact of ergosterol content on acetic and lactic acids toxicity to Saccharomyces cerevisiae.

Organic acid stress often represents a major hurdle in industrial bio-based microbial processes. Organic acids can be released from lignocellulosic feedstocks pretreatment and can also be desirable products obtained by microbial fermentation with applications in different industrial sectors. Yeasts are prominent cell factories. However, the presence of organic acids can compromise yeast metabolism, impairing fermentation performances and limiting the economic feasibility of the processes. Plasma membrane remodeling is deeply involved in yeast tolerance to organic acids, but the detailed mechanisms and potentials of this phenomenon remain largely to be studied and exploited. We investigated the impact of ergosterol on Saccharomyces cerevisiae tolerance against organic acid stress by coupling in vitro and in vivo assays. In the in vitro assay, synthetic lipid vesicles were prepared containing different concentrations of ergosterol. We observed changes in organic acids diffusion through the membrane as a function of ergosterol content. Then, we extended our approach in vivo, engineering S. cerevisiae with the aim of changing the ergosterol content of cells. We focused on ECM22, an important transcription factor, involved in the regulation of ergosterol biosynthesis. The overexpression of ECM22 was sufficient to increase ergosterol levels in S. cerevisiae, resulting in an enhanced tolerance toward lactic acid stress. In this work we propose an in vitro approach, using synthetic lipid vesicles, as a complementary method to be used when studying the impact of the plasma membrane lipid composition on the diffusion of organic acids.

Mold fungi development during the short-term wood-chips storage depending on the storage method

ABSTRACT Wood chips can be stored in a variety of ways and the choice of method can affect their properties. Because mold fungi are common in landfills and in the case of advanced growth the wood material can be classified as low quality, the presented research was carried out to determine the impact of the method of wood chips short-term storage on the development of mold fungi. Chips stored in silo, on unpaved yard, paved yard with and without the roof were analyzed. The genera and species of fungi were identified, their growth was assessed with a visual method, and the concentrations of ergosterol, citric acid, and oxalic acid were determined. The research have shown that from the point of view of mold fungi development, the most advantageous is to store the chips in a silo and the least favorable on the unpaved yard. The effect of roofing of the paved yard probably would be more noticeable during the short-term storage.

Adapting an Ergosterol Extraction Method with Marine Yeasts for the Quantification of Oceanic Fungal Biomass.

Ergosterol has traditionally been used as a proxy to estimate fungal biomass as it is almost exclusively found in fungal lipid membranes. Ergosterol determination has been mostly used for fungal samples from terrestrial, freshwater, salt marsh- and mangrove-dominated environments or to describe fungal degradation of plant matter. In the open ocean, however, the expected concentrations of ergosterol are orders of magnitude lower than in terrestrial or macrophyte-dominated coastal systems. Consequently, the fungal biomass in the open ocean remains largely unknown. Recent evidence based on microscopy and -omics techniques suggests, however, that fungi contribute substantially to the microbial biomass in the oceanic water column, highlighting the need to accurately determine fungal biomass in the open ocean. We performed ergosterol extractions of an oceanic fungal isolate (Rhodotorula sphaerocarpa) with biomass concentrations varying over nine orders of magnitude. While after the initial chloroform-methanol extraction ~87% of the ergosterol was recovered, a second extraction recovered an additional ~10%. Testing this extraction method on samples collected from the open Atlantic Ocean, we successfully determined ergosterol concentrations as low as 0.12 pM. Thus, this highly sensitive method is well suited for measuring fungal biomass from open ocean waters, including deep-sea environments.

The Fungicide Tebuconazole Confounds Concentrations of Molecular Biomarkers Estimating Fungal Biomass.

Due to their ecological importance, fungi are suitable indicator organisms for anthropogenic stress. To estimate fungal biomass, the fungal membrane molecule ergosterol is often quantified as a proxy. Estimates based on ergosterol may, however, be distorted by exposure to demethylase inhibiting (DMI) fungicides, interfering with sterol synthesis. To test this hypothesis, we exposed ten fungal species to the DMI fungicide tebuconazole and measured concentrations of ergosterol and DNA per unit dry mass of the fungal hyphae. The latter served as alternative biomass proxy that is not specifically targeted by tebuconazole. Effects of tebuconazole on ergosterol concentrations were species-specific, while concentrations were on average reduced by 13%. In contrast, DNA concentrations were on average increased by 13%. We demonstrate that DMI fungicides - at close to field relevant levels - can distort fungal biomass estimation, complicating the use of this endpoint for environmental management.

Nitrogen and Ergosterol Concentrations Varied in Live Jack Pine Phloem Following Inoculations With Fungal Associates of Mountain Pine Beetle.

Bark beetles form symbiotic associations with multiple species of fungi that supplement their metabolic needs. However, the relative contributions of each symbiont to the nutrition of bark beetles have been largely unexplored. Thus, we evaluated the ability of three fungal symbionts of mountain pine beetle to concentrate nitrogen and produce ergosterol while infecting phloem of a novel host jack pine. Ergosterol was used as proxy to determine the fungal biomass (hyphal density) in the current study. We inoculated 80 trees in two forest stands with one of the three fungal species or a non-fungal (control) agar. Six weeks later, we collected phloem from the necrotic lesions induced by the fungi, uninfected tissues adjacent to lesions, and non-inoculated control trees. We found that nutritional contributions varied with fungal species. Nitrogen in lesions was higher in trees inoculated with Ophiostoma montium or control trees, relative to Grosmannia clavigera or Leptographium longiclavatum. Furthermore, concentrations of ergosterol were higher in O. montium lesions compared to other tissues or treatments. These results suggest that O. montium differs from G. clavigera and L. longiclavatum in terms of acquiring nitrogen from host tissues and producing ergosterol.

Ergosterol exerts a differential effect on AR-dependent LNCaP and AR-independent DU-145 cancer cells

Androgen-dependent LNCaP and androgen-independent DU-145 cells, were treated with different concentrations of ergosterol (15 µM and 25 µM) and its respective cell viability was measured by MTT bioassay. While ergosterol showed an antiproliferative effect on LNCaP, on DU-145 promoted cell proliferation. This differential effect suggests that the effect of ergosterol might be related to its ability to act as an Androgen Receptor ligand. In silico Molecular Dynamics simulations were performed to analyze the interaction mechanism between androgen receptor and ergosterol, in comparison with natural ligands, 5α-dihydrotestosterone and testosterone. Our model suggests that the binding of androgen receptor with ergosterol is thermodinamically feasible, which is concordant with our experimental results.

Wood processing waste – contamination with microscopic fungi and contents of selected bioactive compounds

Wood processing work stations produce contaminants that affect air quality in plant production facilities. A significant portion of these contaminants consists of spores and hyphae of microscopic fungi. Their presence in respirable and settled dust directly affects the health of the employees working in those facilities. Moreover, microscopic fungi interact with the components of wood, causing its degradation. Thus, several factors affecting the quality of ambient air were analyzed in samples collected from all accessible locations where wood waste is accumulated in the plant. The samples were tested for their concentrations of ergosterol, total phenolics, and antioxidant activity as well as their contents of endogenous wood sterols such as desmosterol, cholesterol, lanosterol, stigmasterol, and beta-sitosterol. The analyses showed that wood waste, despite the varied location and exposure time, promotes the growth of microscopic fungi. Several significant correlations between the analyzed parameters became evident, which made it possible to design the sterol bioconversion mechanism for wood, taking place as a result of the growth of microscopic fungi on the wood material.

Identification of plant genes putatively involved in the perception of fungal ergosterol-squalene.

Trichoderma biocontrol strains establish a complex network of interactions with plants, in which diverse fungal molecules are involved in the recognition of these fungi as nonpathogenic organisms. These molecules act as microbial‐associated molecular patterns that trigger plant responses. Previous studies have reported the importance of ergosterol produced by Trichoderma spp. for the ability of these fungi to induce plant growth and defenses. In addition, squalene, a sterol biosynthetic intermediate, seems to play an important role in these interactions. Here, we analyzed the effect of different concentrations of ergosterol and squalene on tomato (Solanum lycopersicum) growth and on the transcription level of defense‐ and growth‐related genes. We used an RNA‐seq strategy to identify several tomato genes encoding predicted pattern recognition receptor proteins or WRKY transcription factors, both of which are putatively involved in the perception and response to ergosterol and squalene. Finally, an analysis of Arabidopsis thaliana mutants lacking the genes homologous to these tomato candidates led to the identification of a WRKY40 transcription factor that negatively regulates salicylic acid‐related genes and positively regulates ethylene‐ and jasmonate‐related genes in the presence of ergosterol and squalene.

Predicting climate change impacts on maritime Antarctic soils: a space-for-time substitution study

We report a space-for-time substitution study predicting the impacts of climate change on vegetated maritime Antarctic soils. Analyses of soils from under Deschampsia antarctica sampled from three islands along a 2200 km climatic gradient indicated that those from sub-Antarctica had higher moisture, organic matter and carbon (C) concentrations, more depleted δ13C values, lower concentrations of the fungal biomarker ergosterol and higher concentrations of bacterial PLFA biomarkers and plant wax n-alkane biomarkers than those from maritime Antarctica. Shallow soils (2 cm depth) were wetter, and had higher concentrations of organic matter, ergosterol and bacterial PLFAs, than deeper soils (4 cm and 8 cm depths). Correlative analyses indicated that factors associated with climate change (increased soil moisture, C and organic matter concentrations, and depleted δ13C contents) are likely to give rise to increases in Gram negative bacteria, and decreases in Gram positive bacteria and fungi, in maritime Antarctic soils. Bomb-14C analyses indicated that sub-Antarctic soils at all depths contained significant amounts of modern 14C (C fixed from the atmosphere post c. 1955), whereas modern 14C was restricted to depths of 2 cm and 4 cm in maritime Antarctica. The oldest C (c. 1745 years BP) was present in the southernmost soil. The higher nitrogen (N) concentrations and δ15N values recorded in the southernmost soil were attributed to N inputs from bird guano. Based on these analyses, we conclude that 5–8 °C rises in air temperature, together with associated increases in precipitation, are likely to have substantial impacts on maritime Antarctic soils, but that, at the rates of climate warming predicted under moderate greenhouse gas emission scenarios, these impacts are likely to take at least a century to manifest themselves.

A glance into the black box: Novel species-specific quantitative real-time PCR assays to disentangle aquatic hyphomycete community composition

Abstract Aquatic hyphomycetes (AH) are ubiquitous fungi playing a key role in the decomposition of leaf litter in streams. Though their functional performance is modulated by their community composition, this ecological relationship remains poorly investigated due to a lack of suitable methods to identify the biomass-contribution of individual species to AH communities. We, therefore, designed and validated TaqMan® probe-based qPCR assays targeting ten AH species common in temperate regions, allowing detection and quantification of these species within complex communities. In a further step, we compared qPCR-obtained DNA levels to concentrations of the traditional fungal biomass proxy ergosterol. We demonstrate that the qPCR assays are valid for use and that DNA and ergosterol concentrations were significantly positively correlated, suggesting DNA levels as a suitable species-specific biomass proxy. Accordingly, the use of these assays may facilitate multi-species experiments to address major research issues in stress and community ecology including biodiversity-ecosystem functioning relationships.

Intake of ergosterol increases the vitamin D concentrations in serum and liver of mice

Factors that can modify the bioavailability of orally administered vitamin D are not yet widely known. Ergosterol is a common fungal sterol found in food which has a chemical structure comparable to that of vitamin D. This study aimed to investigate the effect of ergosterol on vitamin D metabolism. Therefore, 36 male wild type-mice were randomly subdivided into three groups (n = 12) and received a diet containing 25 μg vitamin D3 and either 0 mg (control), 2 mg or 7 mg ergosterol per kg diet for 6 weeks. To elucidate the impact of ergosterol on hepatic hydroxylation of vitamin D, human hepatoma cells (HepG2) were treated with different concentrations of ergosterol. Concentrations of vitamin D3 and 25-hydroxyvitamin D3 (25(OH)D3) in cells, livers and kidneys of mice and additionally 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) in serum were quantified by LC-MS/MS. The concentration of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) in serum was analyzed by commercially-available enzyme immuno assay. The concentrations of cholesterol and triglycerides were analyzed in livers of mice by photometric assays.Analyses revealed that mice receiving 7 mg/kg ergosterol with their diet had 1.3-, 1.7- and 1.5-times higher concentrations of vitamin D3 in serum, liver and kidney, respectively, than control mice (P < 0.05), whereas no significant effects were observed in mice fed 2 mg/kg ergosterol. The hydroxylation of vitamin D remained unaffected by dietary ergosterol, since the concentration of 25-hydroxyvitamin D3 in serum and tissues and the concentrations of 1,25(OH)2D3 and 24,25(OH)2D3 in serum were not different between the three groups of mice. The lipid concentrations in liver were also not affected by dietary ergosterol. Data from the cell culture studies showed that ergosterol did not influence the conversion of vitamin D3 to 25(OH)D3. To conclude, ergosterol appears to be a modulator of vitamin D3 concentrations in the body of mice, without modulating the hydroxylation of vitamin D3 in liver.

Quantitative profile of phenolic acids and antioxidant activity of wheat grain exposed to stress

In the case of biotic and abiotic stresses, natural defence mechanisms are triggered in plants. The mechanisms are based mainly on enzymatic or non-enzymatic intracellular reactions. Changes in the quantitative profile of 12 phenolic acids, total bound phenolic acids, total free phenolic acids and antioxidant activity in grain of 23 winter wheat genotypes exposed to stress (artificial inoculation with spores of fungi from the genus Fusarium or chemical protection) were analysed. Concentrations of ergosterol confirm significantly higher levels of contamination with microscopic fungi in inoculated wheat samples in comparison to the control and chemically protected samples. The highest concentrations of bound phenolic acids were recorded in inoculated samples while the control and chemically protected samples did not differ significantly. Chromatographic analysis of contents of 12 bound phenolic acids showed that significant differences were found for contents of ferulic, synapic, t-cinnamic and 4-hydroxybenzoic acids in experimental variants. Stepwise discriminatory analysis based on the contents of selected bound phenolic acids antioxidant activity and free phenolic acids led to a complete separation of the investigated populations of the experimental groups. The statistical analysis confirmed that free phenolic acids are the most significant factors in resistance mechanisms as the first line of defence together with their antioxidant action, followed by the synergistic effect of exogenous bound phenolic acids: ferulic, synapic, t-cinnamic and 4-hydroxybenzoic acids.

Vitamin D2 Stability During the Refrigerated Storage of Ultraviolet B-Treated Cultivated Culinary-Medicinal Mushrooms.

The effects of ultraviolet B (UVB) irradiation on the synthesis of vitamin D2 and its stability during refrigerated storage was determined in fresh cultivated culinary-medicinal mushrooms (Agaricus bisporus, Pleurotus ostreatus, and Lentinus edodes) after harvest. The irradiated mushrooms were stored at 4°C for up to 10 days. The concentrations of vitamin D2 and ergosterol were determined using ultrahigh-performance liquid chromatography/tandem mass spectrometry. The cultivated mushrooms not treated with UVB were devoid of vitamin D2. After UVB irradiation, we obtained mushrooms with a large amount of ergocalciferol. A. bisporus showed the lowest vitamin D2 content (3.55 ± 0.11 μg D2/g dry weight); P. ostreatus contained 58.96 ± 1.15 μg D2/g dry weight, and L. edodes contained 29.46 ± 2.21 μg/g dry weight. During storage at 4°C, the amount of vitamin D2 was gradually decreased in P. ostreatus and L. edodes, whereas in A. bisporus vitamin D2 gradually increased until the sixth day, then decreased. Mushrooms exposed to UVB radiation contain a significant amount of vitamin D2 and are therefore an excellent food source of vitamin D.

Assessment of exposure to fungi in the heavily contaminated work environment (a solid waste sorting plant) based on the ergosterol analysis.

This paper reports on the results of the study aimed at application of ergosterol as an quantitative indicator of fungal bioaerosol present in the indoor air in occupational environment heavily contaminated with organic dust as well as its comparison with the culturable method. The study was conducted in the indoor solid waste sorting plant. Using Andersen impactor adapted to 1 plate at the flow rate of 30 l/min, indoor air was sampled in the workers' breathing zone. Ergosterol was sampled using gelatinous filter (1000 l of air) and then analyzed by means of the spectrophotometric method. Fungi were sampled on malt extract agar (MEA) medium (3 replications: 2 l, 7.5 l, 15 l of air) and analyzed by means of the culturable method. Based on ergosterol analyzes, concentration of fungi was calculated. Results were given as the range assuming min. as 5.1 pg ergosterol/spore and max as 1.7 pg ergosterol/spore. The average concentrations of ergosterol in a working room (arithmetic mean (AM), standard deviation (SD); minimum-maximum (min.-max)) were, respectively: 2.16, 0.72; 0.85-2.92 μg/m3; fungi calculated based on ergosterol - 424.1×103-1272.4×103, 140.1×103- 420.4×103, 167×103-1716.5×103 CFU/m3, and culturable fungi - 13×103, 9.7×103, 1.9×103-34×103 CFU/m3). It was revealed that concentrations of calculated fungi were even 2 orders of magnitude higher than culturable fungi. The quantitative assessment of moldiness by means of ergosterol measurement seems to be a reliable indicator for environments heavily contaminated with organic dust, where viable and non-viable fungi are present in high proportions. Based on that result, more restrictive (as compared to a similar assessment carried out by means of the culturable method) hygienic recommendations, especially those related to the use of preventive measures protecting the employees' respiratory tract, should have been undertaken.

Temporal variations of the distribution of water-stable aggregates, microbial biomass and ergosterol in temperate grassland soils with different cultivation histories

In grassland soils, the temporal dynamics in different fractions of soil organic carbon (SOC) is not completely understood. We investigated the temporal variations of water-stable aggregates, microbial biomass C (Cmic) and ergosterol concentrations (as a proxy for fungal biomass) in loamy sandy soils under grassland (north of Kiel, Germany). Samples were taken six times between October 2010 and October 2011 from i) a permanent grassland, ii) an arable plot converted to grassland and iii) a grassland with a one-time tillage operation (both in September 2010). Water-stable aggregate concentrations were determined by a wet-sieving procedure, Cmic concentrations by chloroform-fumigation-extraction and ergosterol concentrations by ethanol extraction. Temporal variation in aggregate distribution of soil samples from the permanent grassland was considerable: the large macroaggregate (>2000μm) concentrations (mean±standard error, n=3) in the surface soil (0–10cm) were highest in October 2011 (666±12gkg−1) and lowest in May (206±49gkg−1). The tillage operation in grassland led to lower concentrations of large macroaggregates, ergosterol and Cmic in the surface soil than in the permanent grassland. The conversion of arable land into grassland did not affect macroaggregate concentrations, despite an increase in Cmic (1.4-fold) and ergosterol concentration (3.3-fold) in the surface soil. Multiple linear regression analysis described the variations in the large macroaggregate concentration satisfactorily (R2=0.60) and indicated that the gravimetric moisture content and the Cmic concentration had positive effects and the pH a negative effect. While ergosterol concentrations (mean±standard error) showed some temporal variations (values for the surface soil in the grassland ranged from 0.5±0.1 to 1.5±0.1mgkg−1), Cmic concentrations varied much less, indicating a higher sensitivity of ergosterol to soil cultivation or changing environmental conditions.

Effect of ergosterol-enriched extracts obtained from Agaricus bisporus on cholesterol absorption using an in vitro digestion model

Ergosterol-enriched extracts obtained from Agaricus bisporus were incorporated into lard, butter and white chocolate as food carriers to study their potential as ingredients with hypocholesterolemic properties to design functional foods. Lard was selected and supplemented with several concentrations of cholesterol and ergosterol, beta-sitosterol (as control) and two types of ergosterol-enriched extracts obtained by supercritical fluid extraction (SFE) and accelerated solvent extraction (ASE). The prepared food combinations were submitted to a specific in vitro digestion model, their dietary mixed micelle (DMM) fractions isolated and administrated to Caco2 cell cultures. Ergosterol was as effective as some phytosterols such as sitosterol, in the displacement of cholesterol from the DMMs when applied to the food matrix in a ratio (1:2, w/w, cholesterol:sterol). The ASE extract was not effective but addition of the SFE extract provoked a 67% reduction in the DMM cholesterol content. Lard supplementation with fungal beta-glucans able to bind bile-acids enhanced the cholesterol displacement from the DMMs induced by the sterols although it also reduced ergosterol incorporation. Fungal sterols also impaired proper cholesterol transport through Caco2 monolayers.

Overproduction of geranylgeraniol in Coprinopsis cinerea by the expression of geranylgeranyl diphosphate synthase gene.

(E, E, E)-Geranylgeraniol (GGOH) is a valuable ingredient of many perfumes and a valuable precursor for synthesizing pharmaceuticals. In an attempt to increase the GGOH concentration in Coprinopsis cinerea, we demonstrated that the expression of geranylgeranyl diphosphate synthase (ggpps) gene isolated from Taxus x media could promote GGOH production. Furthermore, the concentrations of squalene and ergosterol were measured in the engineered strains. Expectedly, significant decreases of squalene and ergosterol levels were observed in those strains transformed with ggpps gene. This could be explained by the partial redirection of metabolic flux from squalene to GGOH, whose biosynthesis competes for the same precursor with squalene. This work suggested that the expression of ggpps in higher fungi was an effective method for bio-production of GGOH.