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The consequence of the massive increase in population in recent years is the enormous production of mainly industrial waste. The effort to minimize these waste products is, therefore, no longer sufficient. Biotechnologists, therefore, started looking for ways to not only reuse these waste products, but also to valorise them. This work focuses on the biotechnological use and processing of waste oils/fats and waste glycerol by carotenogenic yeasts of the genus Rhodotorula and Sporidiobolus. The results of this work show that the selected yeast strains are able to process waste glycerol as well as some oils and fats in a circular economy model and, moreover, are resistant to potential antimicrobial compounds present in the medium. The best-growing strains, Rhodotorula toruloides CCY 062-002-004 and Rhodotorula kratochvilovae CCY 020-002-026, were selected for fed-batch cultivation in a laboratory bioreactor in a medium containing a mixture of coffee oil and waste glycerol. The results show that both strains were able to produce more than 18 g of biomass per litre of media with a high content of carotenoids (10.757 +/- 1.007 mg/g of CDW in R. kratochvilovae and 10.514 +/- 1.520 mg/g of CDW in R. toruloides, respectively). The overall results prove that combining different waste substrates is a promising option for producing yeast biomass enriched with carotenoids, lipids, and beta-glucans.

The consequence of the massive increase in population in recent years is the enormous production of mainly industrial waste. The effort to minimize these waste products is, therefore, no longer sufficient. Biotechnologists, therefore, started looking for ways to not only reuse these waste products, but also to valorise them. This work focuses on the biotechnological use and processing of waste oils/fats and waste glycerol by carotenogenic yeasts of the genus Rhodotorula and Sporidiobolus. The results of this work show that the selected yeast strains are able to process waste glycerol as well as some oils and fats in a circular economy model and, moreover, are resistant to potential antimicrobial compounds present in the medium. The best-growing strains, Rhodotorula toruloides CCY 062-002-004 and Rhodotorula kratochvilovae CCY 020-002-026, were selected for fed-batch cultivation in a laboratory bioreactor in a medium containing a mixture of coffee oil and waste glycerol. The results show that both strains were able to produce more than 18 g of biomass per litre of media with a high content of carotenoids (10.757 +/- 1.007 mg/g of CDW in R. kratochvilovae and 10.514 +/- 1.520 mg/g of CDW in R. toruloides, respectively). The overall results prove that combining different waste substrates is a promising option for producing yeast biomass enriched with carotenoids, lipids, and beta-glucans.

The consequence of the massive increase in population in recent years is the enormous production of mainly industrial waste. The effort to minimize these waste products is, therefore, no longer sufficient. Biotechnologists, therefore, started looking for ways to not only reuse these waste products, but also to valorise them. This work focuses on the biotechnological use and processing of waste oils/fats and waste glycerol by carotenogenic yeasts of the genus Rhodotorula and Sporidiobolus. The results of this work show that the selected yeast strains are able to process waste glycerol as well as some oils and fats in a circular economy model and, moreover, are resistant to potential antimicrobial compounds present in the medium. The best-growing strains, Rhodotorula toruloides CCY 062-002-004 and Rhodotorula kratochvilovae CCY 020-002-026, were selected for fed-batch cultivation in a laboratory bioreactor in a medium containing a mixture of coffee oil and waste glycerol. The results show that both strains were able to produce more than 18 g of biomass per litre of media with a high content of carotenoids (10.757 +/- 1.007 mg/g of CDW in R. kratochvilovae and 10.514 +/- 1.520 mg/g of CDW in R. toruloides, respectively). The overall results prove that combining different waste substrates is a promising option for producing yeast biomass enriched with carotenoids, lipids, and beta-glucans.

Anti-cancer T-cell responses are often halted due to the immune-suppressive micro-environment, in part related to tumor-associated macrophages. In the current study, we assessed indigestible β-glucans (oatβG, curdlan, grifolan, schizophyllan, lentinan, yeast whole glucan particles (yWGP), zymosan and two additional yeast-derived β-glucans a and b) for their physicochemical properties as well as their effects on the plasticity of human monocyte-derived macrophages that were polarized with IL-4 to immune-suppressive macrophages. Beta-glucans were LPS/LTA free, and tested for solubility, molecular masses, protein and monosaccharide contents. Curdlan, yeast-b and zymosan re-polarized M(IL-4) macrophages towards an M1-like phenotype, in particular showing enhanced gene expression of CCR7, ICAM1 and CD80, and secretion of TNF-α and IL-6. Notably, differential gene expression, pathway analysis as well as protein expressions demonstrated that M(IL-4) macrophages treated with curdlan, yeast-b or zymosan demonstrated enhanced production of chemo-attractants, such as CCL3, CCL4, and CXCL8, which contribute to recruitment of monocytes and neutrophils. The secretion of chemo-attractants was confirmed when using patient-derived melanoma-infiltrating immune cells. Taken together, the bacterial-derived curdlan as well as the yeast-derived β-glucans yeast-b and zymosan have the unique ability to preferentially skew macrophages towards a chemo-attractant-producing phenotype that may aid in anti-cancer immune responses.

The epithelium-associated cytokine thymic stromal lymphopoietin (TSLP) can induce OX40L and CCL17 expression by myeloid dendritic cells (mDCs), which contributes to aberrant Th2-type immune responses. Herein, we report that such TSLP-induced Th2-type immune response can be effectively controlled by Dectin-1, a C-type lectin receptor expressed by mDCs. Dectin-1 stimulation induced STAT3 activation and decreased the transcriptional activity of p50-RelB, both of which resulted in reduced OX40L expression on TSLP-activated mDCs. Dectin-1 stimulation also suppressed TSLP-induced STAT6 activation, resulting in decreased expression of the Th2 chemoattractant CCL17. We further demonstrated that Dectin-1 activation was capable of suppressing ragweed allergen (Amb a 1)-specific Th2-type T cell response in allergy patients ex vivo and house dust mite allergen (Der p 1)-specific IgE response in non-human primates in vivo. Collectively, this study provides a molecular explanation of Dectin-1-mediated suppression of Th2-type inflammatory responses and suggests Dectin-1 as a target for controlling Th2-type inflammation.

Background(1-3)-b-D-glucan (BDG) is a fungal cell wall component and, in the absence of invasive fungal infection, a novel biomarker for microbial translocation of endogenous fungal products from the gastrointestinal tract into systemic circulation. However, its value as a marker of fungal translocation is limited by a concern that plant BDG-rich food influences blood BDG levels.MethodsWe conducted a pilot clinical trial to evaluate the impact of a standardised oral BDG challenge on blood BDG levels in participants with and without elevated microbial translocation. We enrolled 14 participants including 8 with HIV infection, 2 with advanced liver cirrhosis, and 4 healthy controls. After obtaining a baseline blood sample, participants received a standardised milkshake containing high levels of BDG followed by serial blood samples up to 8 hours after intake.ResultsThe standardised oral BDG challenge approach did not change the blood BDG levels over time in all participants. We found consistently elevated blood BDG levels in one participant with advanced liver cirrhosis and a single person with HIV with a low CD4 count of 201 cells/mm3 .ConclusionOur findings indicate that BDG blood levels were not influenced by plant origin BDG-rich nutrition in PWH, people with advanced liver cirrhosis, or healthy controls. Future studies are needed to analyse gut mycobiota populations in individuals with elevated blood BDG levels.

Beta (beta)-glucans are polysaccharides composed of D-glucose monomers. Nowadays, beta-glucans are gaining attention due to their attractive immunomodulatory biological activities, which can be utilized in pharmaceutical or food supplementation industries. Some carotenogenicBasidiomycetesyeasts, previously explored for lipid and carotenoid coproduction, could potentially coproduce a significant amount of beta-glucans. In the present study, we screened elevenBasidiomycetesfor the coproduction of lipids and beta-glucans. We examined the effect of four different C/N ratios and eight different osmolarity conditions on the coproduction of lipids and beta-glucans. A high-throughput screening approach employing microcultivation in microtiter plates, Fourier Transform Infrared (FTIR) spectroscopy and reference analysis was utilized in the study. Yeast strainsC. infirmominiatumCCY 17-18-4 andR. kratochvilovaeCCY 20-2-26 were identified as the best coproducers of lipids and beta-glucans. In addition,C. infirmominiatumCCY 17-18-4,R. kratochvilovaeCCY 20-2-26 andP. rhodozymaCCY 77-1-1 were identified as the best alternative producers of beta-glucans. Increased C/N ratio led to increased biomass, lipid and beta-glucans production for several yeast strains. Increased osmolarity had a negative effect on biomass and lipid production while the beta-glucan production was positively affected.

Beta (beta)-glucans are polysaccharides composed of D-glucose monomers. Nowadays, beta-glucans are gaining attention due to their attractive immunomodulatory biological activities, which can be utilized in pharmaceutical or food supplementation industries. Some carotenogenicBasidiomycetesyeasts, previously explored for lipid and carotenoid coproduction, could potentially coproduce a significant amount of beta-glucans. In the present study, we screened elevenBasidiomycetesfor the coproduction of lipids and beta-glucans. We examined the effect of four different C/N ratios and eight different osmolarity conditions on the coproduction of lipids and beta-glucans. A high-throughput screening approach employing microcultivation in microtiter plates, Fourier Transform Infrared (FTIR) spectroscopy and reference analysis was utilized in the study. Yeast strainsC. infirmominiatumCCY 17-18-4 andR. kratochvilovaeCCY 20-2-26 were identified as the best coproducers of lipids and beta-glucans. In addition,C. infirmominiatumCCY 17-18-4,R. kratochvilovaeCCY 20-2-26 andP. rhodozymaCCY 77-1-1 were identified as the best alternative producers of beta-glucans. Increased C/N ratio led to increased biomass, lipid and beta-glucans production for several yeast strains. Increased osmolarity had a negative effect on biomass and lipid production while the beta-glucan production was positively affected.

Beta (beta)-glucans are polysaccharides composed of D-glucose monomers. Nowadays, beta-glucans are gaining attention due to their attractive immunomodulatory biological activities, which can be utilized in pharmaceutical or food supplementation industries. Some carotenogenicBasidiomycetesyeasts, previously explored for lipid and carotenoid coproduction, could potentially coproduce a significant amount of beta-glucans. In the present study, we screened elevenBasidiomycetesfor the coproduction of lipids and beta-glucans. We examined the effect of four different C/N ratios and eight different osmolarity conditions on the coproduction of lipids and beta-glucans. A high-throughput screening approach employing microcultivation in microtiter plates, Fourier Transform Infrared (FTIR) spectroscopy and reference analysis was utilized in the study. Yeast strainsC. infirmominiatumCCY 17-18-4 andR. kratochvilovaeCCY 20-2-26 were identified as the best coproducers of lipids and beta-glucans. In addition,C. infirmominiatumCCY 17-18-4,R. kratochvilovaeCCY 20-2-26 andP. rhodozymaCCY 77-1-1 were identified as the best alternative producers of beta-glucans. Increased C/N ratio led to increased biomass, lipid and beta-glucans production for several yeast strains. Increased osmolarity had a negative effect on biomass and lipid production while the beta-glucan production was positively affected.

Systematic reproduction and distribution, duplication of any material in this paper for a fee or for commercial purposes, or modifications of the content of this paper are prohibited., [EN] The present study evaluates the effect of dietary barley, based on its potential stress-relieving properties, on rainbow trout under acute stress challenge (hypoxia and crowding) and their recovery. Diets were formulated containing increasing barley concentrations (0, 4, 8, 16, 32%). Cortisol on plasma and fin, glucose and lactate plasma levels and malondialdehyde (MDA) in muscle were determined under normoxia before the stress test (basal levels), 30 min after the acute stress challenge and also during normoxia recovery (6 and 12 h after the stress). Results showed that at basal levels the inclusion of barley had no influence on cortisol, glucose nor on lactate values. After 30 min from the stress challenge, there was a significant increase in cortisol, glucose and lactate concentration in fish of all groups. Plasma cortisol showed the lowest levels in fish fed with diets at a medium (8%) of barley concentration and returned to basal levels 6 h after the stress stimulus with no differences between diets. Glucose values showed a less clear tendency 30 min after the stress challenge with lower levels in the control group, fish fed with 8% and 32% of barley in the diets and returned to basal levels in almost all the groups only 12 h after the stress challenge. Lactate showed the same trend as with glucose after the stress challenge but it returned to basal levels in 6 h. Interestingly, there was a significant decrease of lipid oxidation (MDA) in muscle soon after the stress test of fish fed with the highest barley levels. The present results suggest a potential positive effect of dietary barley on trout stress response.

Systematic reproduction and distribution, duplication of any material in this paper for a fee or for commercial purposes, or modifications of the content of this paper are prohibited., [EN] The present study evaluates the effect of dietary barley, based on its potential stress-relieving properties, on rainbow trout under acute stress challenge (hypoxia and crowding) and their recovery. Diets were formulated containing increasing barley concentrations (0, 4, 8, 16, 32%). Cortisol on plasma and fin, glucose and lactate plasma levels and malondialdehyde (MDA) in muscle were determined under normoxia before the stress test (basal levels), 30 min after the acute stress challenge and also during normoxia recovery (6 and 12 h after the stress). Results showed that at basal levels the inclusion of barley had no influence on cortisol, glucose nor on lactate values. After 30 min from the stress challenge, there was a significant increase in cortisol, glucose and lactate concentration in fish of all groups. Plasma cortisol showed the lowest levels in fish fed with diets at a medium (8%) of barley concentration and returned to basal levels 6 h after the stress stimulus with no differences between diets. Glucose values showed a less clear tendency 30 min after the stress challenge with lower levels in the control group, fish fed with 8% and 32% of barley in the diets and returned to basal levels in almost all the groups only 12 h after the stress challenge. Lactate showed the same trend as with glucose after the stress challenge but it returned to basal levels in 6 h. Interestingly, there was a significant decrease of lipid oxidation (MDA) in muscle soon after the stress test of fish fed with the highest barley levels. The present results suggest a potential positive effect of dietary barley on trout stress response.

[EN] This study investigated the effects of the incorporation of barley instead of wheat to study fish growth and biometrics parameters, body composition, retention efficiency and liver histology of rainbow trout Oncorhynchus mykiss. Experimental diets were used containing different barley concentrations (0-16%) randomly assigned to four groups per treatment. Results showed a significant change on liver histological parameters. Liver cell vacuolization was significantly lower on fish fed with 16B diets compared to control diet and revealed some degeneration. Those fish also showed more regular hepatocytes than in control diet.

[EN] Barley concentrations ranging from 0 to 32% (0B, 40B, 80B, 160B, and 319B) were incorporated into rainbow trout, Oncorhynchus mykiss (Walbaum) diets. The experiment started with an initial average fish weight of 127.72 +/- 5.65 g and finished when they reached commercial weight (final weight between 312 and 330 g) after 84 days. The inclusion of barley in the diets did not show a significant effect on growth and biometric parameters, fat and carbohydrate digestibilities; however, protein digestibility decreased significantly with the incorporation of barley on diets. Glucose levels increased significantly with barley concentration in the diet, and lactate and cortisol levels were also significantly affected after a stress period regardless of the diet. Meat quality was influenced as well by barley concentration. Lower water activity values and an enhancement in textural and color properties were observed in fish fed with the diet containing the highest barley concentration. Trout fed feed with higher concentrations of barley (160B) showed lower lipid oxidation levels than those fed with lower concentrations (control and 40B). The sensory panel found that fish fed with diets higher than 8% in barley content (80B) exhibited a brighter red color in the gills and a better texture; also, meat color became redder with a higher barley inclusion (160B and 319B), being all these sensory parameters correlated with fish freshness. Thus, results indicate that barley can be substituted for wheat fraction without any detrimental effect on production efficiency and enhancing fish quality.

Blood citrulline and intestinal fatty acid binding protein were determined as biomarkers for intestinal mucositis. Biomarker levels were correlated with corresponding serum 1,3-beta-D-glucan levels in 56 samples obtained from 33 cases with underlying hematological malignancies receiving induction chemotherapy. No correlation between biomarkers of intestinal mucositis and BDG levels was observed. (This study has been registered at ClinicalTrials.gov under registration no. NCT01576653.).

Elevated total and low density lipoprotein (LDL) cholesterol levels are considered major risk factors for coronary heart disease. Oat is a cereal that is a good source of proteins, fat, minerals and vitamins, as well as soluble fiber, including beta-glucans. Due to the physico- chemical characteristics of the beta-glucan, it has been proposed that oats may help reduce blood cholesterol levels and help control postprandialplasma glucose and insulin response. A large body of clinical studies suggests that the consumption of at least 3 g per day of oat beta-glucan, as part of a diet low in saturated fat and cholesterol, may reduce the risk of coronary heart disease, which has been approved by several regulatory agencies, as the Food and Drug Administration in the USA and the European Food Safety Authority in Europe., Las cifras sanguíneas de colesterol total y de lipoproteínas de baja densidad (LDL) elevadas se consideran como uno de los más importantes factores de riesgo de enfermedad coronaria. La avena es un cereal rico en proteínas, lípidos, vitaminas, minerales y fibra soluble, entre las que se incluye el beta-glucano. Debido a las características físico-químicas del beta-glucano, se ha propuesto que la avena pudiera contribuir a disminuir las cifras de colesterol sanguíneo, así como a controlar la glucosa plasmática postprandial y la respuesta insulínica. Numerosos estudios señalan que el consumo de un mínimo de 3 g/día de beta-glucano de forma regular, como parte de una dieta con un bajo contenido en grasa saturada y colesterol, puede contribuir a disminuir el riesgo de enfermedad coronaria, tal y como han aprobado diversas agencias reguladoras, como la Food and Drug Administration de Estados Unidos o la Agencia Europea de Seguridad Alimentaria.

The Ascomycete Cordyceps militaris, an entomopathogenic fungus, is one of the most important traditional Chinese medicines. Studies related to its pharmacological properties suggest that this mushroom can exert interesting biological activities. Aqueous (CW and HW) and alkaline (K5) extracts containing polysaccharides were prepared from this mushroom, and a ß-D-glucan was purified. This polymer was analysed by GC-MS and NMR spectrometry, showing a linear chain composed of ß-D-Glcp (1¿3)-linked. The six main signals in the 13C-NMR spectrum were assigned by comparison to reported data. The aqueous (CW, HW) extracts stimulated the expression of IL-1ß, TNF-a, and COX-2 by THP-1 macrophages, while the alkaline (K5) extract did not show any effect. However, when the extracts were added to the cells in the presence of LPS, K5 showed the highest inhibition of the pro-inflammatory genes expression. This inhibitory effect was also observed for the purified ß-(1¿3)-D-glucan, that seems to be the most potent anti-inflammatory compound present in the polysaccharide extracts of C. militaris. In vivo, ß-(1¿3)-D-glucan also inhibited significantly the inflammatory phase of formalin-induced nociceptive response, and, in addition, it reduced the migration of total leukocytes but not the neutrophils induced by LPS. In conclusion, this study clearly demonstrates the anti-inflammatory effect of ß-(1¿3)-D-glucan.

Following an application from Leiber GmbH, submitted for authorisation of a health claim pursuant to Article 13(5) of Regulation (EC) No 1924/2006 via the Competent Authority of Germany, the Panel on Dietetic Products, Nutrition and Allergies (NDA) was asked to deliver an opinion on the scientific substantiation of a health claim related to Yestimun® and defence against pathogens in the upper respiratory tract. The food that is the subject of the health claim, Yestimun®, which consists of (1,3)-(1,6)-β-D-glucans from brewer’s yeast cell wall, is sufficiently characterised. The claimed effect, defence against pathogens in the upper respiratory tract, is a beneficial physiological effect. No human intervention studies from which conclusions could be drawn for the scientific substantiation of the claim were provided by the applicant. The Panel concludes that a cause and effect relationship has not been established between the consumption of Yestimun® ((1,3)-(1,6)-β-D-glucans from brewer’s yeast cell wall) and defence against pathogens in the upper respiratory tract. © European Food Safety Authority, 2013

We aimed at evaluating whether beta-glucans could improve low-density lipoprotein (LDL) cholesterol levels and/or glycemic control in patients with myasthenia gravis (MG), in whom statins usually have muscle-related side effects. Fifty-nine MG patients participated in the study and received a daily dietary supplement of 3 g of beta-glucans during 8 weeks. Body mass index (BMI) and blood sample analysis of lipid and glucose status were performed before and after 8 weeks intake of beta-glucans. In the 52 patients who completed the study, there was a significant reduction in total cholesterol, LDL, ApoA1 and ApoB (all P<0.003). However, glycemic control and BMI were unaltered. The present study indicates that 8 weeks daily intake of 3 g of beta-glucans significantly reduces total cholesterol, LDL, ApoA1 and ApoB in MG patients. beta-glucans may therefore be of value in improving lipid status in MG patients, without the muscle-related side effects accompanied by statins.

Background: The differentiation resp. death of human monocytic THP-1 cells induced by polysaccharide extracts of the medicinal mushrooms Phellinus linteus, Agaricus bisporus and Agaricus brasiliensis have been studied. This study aims to identify leads for the causal effects of these mushroom components on cell differentiation and death. Methods: THP-1 cells were treated with different polysaccharide extracts of mushrooms and controls. Morphological effects were observed by light microscopy. Flow cytometry was applied to follow the cell differentiation by cell cycle shifts after staining with propidium iodide, changes of mitochondrial membrane potential after incubation with JC-1, and occurrence of intracellular reactive oxygen species after incubation with hydroethidine. Principal component analysis of the data was performed to evaluate the cellular effects of the different treatments. Results: P. linteus polysaccharide extracts induced dose-dependent apoptosis of THP-1 cells within 24 h, while A. bisporus and A. brasiliensis polysaccharide extracts caused differentiation into macrophages. A pure P. linteus polysaccharide had no effect. Apoptosis was inhibited by preincubating THP-1 cells with human serum. The principal component analysis revealed that P. linteus, A. bisporus and A. brasiliensis polysaccharide extracts increased reactive oxygen species production. Both A. bisporus and A. brasiliensis polysaccharide extracts decreased the mitochondrial membrane potential, while this was increased by P. linteus polysaccharide extracts. Conclusions: P. linteus polysaccharide extracts caused apoptosis of THP-1 monocytes while A. bisporus and A. brasiliensis polysaccharide extracts caused these cells to differentiate into macrophages. The protective effects of human serum suggested that P. linteus polysaccharide extract induced apoptosis by extrinsic pathway, i.e. by binding to the TRAIL receptor. The mitochondrial membrane potential together with reactive oxygen specie

Probiotics, prebiotics and synbiotics are frequently-used components for the elaboration of functional food. Currently, most of the commercialized probiotics are limited to a few strains of the genera Bifidobacteria, Lactobacillus and Streptococcus, most of which produce exopolysaccharides (EPS). This suggests that the beneficial properties of these microorganisms may be related to the biological activities of these biopolymers. In this work we report that a 2-substituted-(1,3)-beta-D-glucan of non-dairy bacterial origin has a prebiotic effect on three probiotic strains. Moreover, the presence of this beta-D-glucan potentiates in vitro adhesion of the probiotic Lactobacillus plantarum WCFS1 to human intestinal epithelial cells.

Probiotics, prebiotics and synbiotics are frequently-used components for the elaboration of functional food. Currently, most of the commercialized probiotics are limited to a few strains of the genera Bifidobacteria, Lactobacillus and Streptococcus, most of which produce exopolysaccharides (EPS). This suggests that the beneficial properties of these microorganisms may be related to the biological activities of these biopolymers. In this work we report that a 2-substituted-(1,3)-beta-D-glucan of non-dairy bacterial origin has a prebiotic effect on three probiotic strains. Moreover, the presence of this beta-D-glucan potentiates in vitro adhesion of the probiotic Lactobacillus plantarum WCFS1 to human intestinal epithelial cells.

Probiotics, prebiotics and synbiotics are frequently-used components for the elaboration of functional food. Currently, most of the commercialized probiotics are limited to a few strains of the genera Bifidobacteria, Lactobacillus and Streptococcus, most of which produce exopolysaccharides (EPS). This suggests that the beneficial properties of these microorganisms may be related to the biological activities of these biopolymers. In this work we report that a 2-substituted-(1,3)-beta-D-glucan of non-dairy bacterial origin has a prebiotic effect on three probiotic strains. Moreover, the presence of this beta-D-glucan potentiates in vitro adhesion of the probiotic Lactobacillus plantarum WCFS1 to human intestinal epithelial cells.

BACKGROUND: Irritable bowel syndrome (IBS) is a very common functional gastrointestinal (GI). Diagnosis of IBS is based on the fulfilment of the Rome III criteria. Common GI symptoms are lower abdominal pain, bloating and disturbed defecation, such as urgent diarrhoea and/or episodes of chronic constipation. Many agents have been employed in the management of IBS, although only few have been demonstrated to show a relevant efficacy. AIM: To evaluate the effectiveness of the administration of a mixture of beta-glucan, inositol and digestive enzymes (Biointo) in improving GI symptoms in patients affected by IBS. PATIENTS AND METHODS: 50 IBS patients (20 males, 30 females; mean age 51 +/- 19) were treated with Biointo (group A) while another group consisting of 40 IBS patients (15 males, 25 females; mean age 50 +/- 18) did not receive any therapy (group B). RESULTS: Biointol administration improved significantly bloating, flatulence and abdominal pain, with a slight increasing of urgency for bowel movements. On the contrary, Biointol did not show any significant effect on the other IBS symptoms. CONCLUSIONS: Currently, only few agents used in the management of IBS have been proven to be effective. Biointol administration has shown to improve some IBS symptoms, such as bloating, flatulence and abdominal pain, all connected to the presence of gas inside the intestinal lumen.

Background Mushroom polysaccharides have traditionally been used for the prevention and treatment of a multitude of disorders like infectious illnesses, cancers and various autoimmune diseases. Crude mushroom extracts have been tested without detailed chemical analyses of its polysaccharide content. For the present study we decided to chemically determine the carbohydrate composition of semi-purified extracts from 2 closely related and well known basidiomycete species, i.e. Agaricus bisporus and A. brasiliensis and to study their effects on the innate immune system, in particular on the in vitro induction of pro-inflammatory cytokines, using THP-1 cells. Methods Mushroom polysaccharide extracts were prepared by hot water extraction and precipitation with ethanol. Their composition was analyzed by GC-MS and NMR spectroscopy. PMA activated THP-1 cells were treated with the extracts under different conditions and the production of pro-inflammatory cytokines was evaluated by qPCR. Results Semi-purified polysaccharide extracts of A. bisporus and A. brasiliensis (= blazei) were found to contain (1¿6),(1¿4)-linked a-glucan, (1¿6)-linked ß-glucan, and mannogalactan. Their proportions were determined by integration of 1H-NMR signs, and were considerably different for the two species. A. brasiliensis showed a higher content of ß-glucan, while A. bisporus presented mannogalactan as its main polysaccharide. The extracts induced a comparable increase of transcription of the pro-inflammatory cytokine genes IL-1ß and TNF-a as well as of COX-2 in PMA differentiated THP-1 cells. Pro-inflammatory effects of bacterial LPS in this assay could be reduced significantly by the simultaneous addition of A. brasiliensis extract. Conclusions The polysaccharide preparations from the closely related species A. bisporus and A. brasiliensis show major differences in composition: A. bisporus shows high mannogalactan content whereas A. brasiliensis has mostly ß-glucan. Semi-purified polysaccharide

Cílem práce bylo popsat taxonomii, fyziologii a chemické složení hub - zvláště se zaměřením na oddělení Basidiomycetes. Práce se také zabývá současnými poznatky o výskytu biologicky aktivních látek v houbách - zejména beta-glukanů. V experimentální části byly u plodnic různých druhů Basidiomycet stanoveny vybrané chemické vlastnosti, a to obsahy sušiny, popelovin, fosforu a rtuti. Část vzorků byla pěstována v domácích podmínkách, část zakoupena v obchodní síti a část nasbírána ve dvou různých lokalitách Valašska., This master thesis is focused on taxonomy, physiology and chemical structure of mushroom - especially aim to division of Basidiomycetes. Attention was payed to present information about occurrence of biological active substances contained in mushrooms - especially beta-glucans. In experimental part were in fruiting bodies of different types of Basidiomycetes analysed chosen chemical properties - contens of dry matter, mineral matter, phosphorus and mercury. Part of saples were grown at home conditions, part was bought and part was picked at two different localities of Wallachia., Ústav potravinářského inženýrství, obhájeno

Cílem práce bylo charakterizovat vybrané sacharidy. Největší pozornost byla věnována beta-glukanům a vláknině potravy. U beta-glukanů byly zjištěny stimulační účinky na imunitní systém člověka. Význam vlákniny ve výživě člověka je nezastupitelný., The objective of this work is to characterize choice descriptions of sugars. The biggest attention was payed to beta-glucans and dietary fibre. Beta-glucans have stimulus effects on im-mune system of human. Meaning of dietray fibre in nutrition of human is not fungible., Ústav potravinářského inženýrství, obhájeno

Epidemiological studies have shown an inverse relation between a whole grain consumption and risk of type-2 diabetes and cardiovascular disease. One tentative mechanism relates to colonic metabolism of indigestible carbohydrates. In a previous study, we reported a positive relation between colonic fermentation and improved glucose tolerance. This work can be seen as an extension of that study, focusing on the tentative role of specific colonic metabolites, i.e. SCFA. Plasma concentrations of acetate, propionate, and butyrate were determined in the morning in healthy participants (5 women and 10 men, mean ± SD: 25.9 ± 3.2 y, BMI <25) following 8 different cereal-based evening meals (50 g available starch) varying in content of indigestible carbohydrates. Each participant consumed all test meals in a random order on separate evenings. At a standardized breakfast following evening test meals, the postprandial glucose response (incremental area under the curve, 0-120 min) was inversely related to plasma butyrate (r = -0.26; P <0.01) and acetate (r = -0.20; P <0.05) concentrations. Evening meals composed of high-amylose barley kernels or high-ß-glucan barley kernels resulted in higher plasma butyrate concentrations the following morning compared with an evening meal with white wheat bread (P <0.05). The results support the view that cereal products rich in indigestible carbohydrates may improve glucose tolerance through a mechanism involving colonic fermentation and generation of SCFA, where in particular butyric acid may be involved. This mechanism may be one explanation by which whole grain is protective against type 2 diabetes and cardiovascular disease.

An a-glucan was isolated from the culinary medicinal mushroom A. bisporus by hot water extraction, ethanol precipitation and DEAE-cellulose chromatography. The resulting material showed a single HMW peak excluded from a Sephadex G50 column that could completely be degraded by a-amylase treatment. After heating in 1% SDS a small additional peak of low MW eluted from the G50 column. The monosaccharide composition of the main peak was evaluated by HPLC, and was found to consist of a majority of glucose (97.6%), and a minor proportion of galactose (2.4%). Methylation analysis and degradation by a-amylase indicated the presence of an a-glucan with a main chain consisting of (1®4)-linked units, substituted at O-6 by a-D-glucopyranose single-units in the relation 1:8. Mono- (13C-, 1H-NMR) and bidimensional [1H (obs.),13C-HSQC] spectroscopy analysis confirmed the a-configuration of the Glcp residues by low frequency resonances of C-1 at d 100.6, 100.2, and 98.8 ppm and H-1 high field ones at d 5.06, 5.11, and 4.74 ppm. The DEPT-13C-NMR allowed assigning the non-substituted and O-substituted –CH2 signals at d 60.3/60.8 and 66.2 ppm, respectively. Other assignments were attributed to C-2, C-3, C-4, C-5 and C-6 of the non-reducing ends at d 71.8; 72.8; 70.0; 71.3 and 60.3/60.8 ppm, respectively. The minor proportion of galactose that was demonstrated was probably derived from a complex between the a-glucan and a low molecular weight galactan

Epidemiological studies have shown an inverse relation between a whole grain consumption and risk of type-2 diabetes and cardiovascular disease. One tentative mechanism relates to colonic metabolism of indigestible carbohydrates. In a previous study, we reported a positive relation between colonic fermentation and improved glucose tolerance. This work can be seen as an extension of that study, focusing on the tentative role of specific colonic metabolites, i.e. SCFA. Plasma concentrations of acetate, propionate, and butyrate were determined in the morning in healthy participants (5 women and 10 men, mean ± SD: 25.9 ± 3.2 y, BMI <25) following 8 different cereal-based evening meals (50 g available starch) varying in content of indigestible carbohydrates. Each participant consumed all test meals in a random order on separate evenings. At a standardized breakfast following evening test meals, the postprandial glucose response (incremental area under the curve, 0-120 min) was inversely related to plasma butyrate (r = -0.26; P <0.01) and acetate (r = -0.20; P <0.05) concentrations. Evening meals composed of high-amylose barley kernels or high-ß-glucan barley kernels resulted in higher plasma butyrate concentrations the following morning compared with an evening meal with white wheat bread (P <0.05). The results support the view that cereal products rich in indigestible carbohydrates may improve glucose tolerance through a mechanism involving colonic fermentation and generation of SCFA, where in particular butyric acid may be involved. This mechanism may be one explanation by which whole grain is protective against type 2 diabetes and cardiovascular disease.

Microbial biofilms are a dominant feature of many human infections. However, developing effective strategies for controlling biofilms requires an understanding of the underlying biology well beyond what currently exists. Using a novel strategy, we have induced formation of a robust biofilm in Escherichia coli by utilizing an exogenous source of poly-N-acetylglucosamine (PNAG) polymer, a major virulence factor of many pathogens. Through microarray profiling of competitive selections, carried out in both transposon insertion and over-expression libraries, we have revealed the genetic basis of PNAG-based biofilm formation. Our observations reveal the dominance of electrostatic interactions between PNAG and surface structures such as lipopolysaccharides. We show that regulatory modulation of these surface structures has significant impact on biofilm formation behavior of the cell. Furthermore, the majority of clinical isolates which produced PNAG also showed the capacity to respond to the exogenously produced version of the polymer.

Barley is a desirable food ingredient, with health benefits provided by a p-glucan fibre fraction. A hull-less barley flour and flakes were incorporated into white and wholegrain wheat bread in quantities usually applied in practice. The breads were evaluated for nutritional composition and sensory properties and compared to standard products as controls. The supplemented breads were high in fibre, zinc and selenium content. It was estimated that a 300-gram daily portion of such breads could meet up to 40% of dietary recommended intakes for selenium and 70-75% of recommended daily values for beta-glucan. Regarding sensory quality, the only significant differences (p lt 0.05) were higher taste and lower volume in the white supplemented breads and lower crumb elasticity in the white bread made with barley flour. Hull-less barley can substantially contribute to an adequate intake of selenium and beta-glucan. In addition, supplemented breads were not found to pose a significant risk, with regard to excessive intakes of heavy elements (Pb, Cd, As).

Vedle chitinu obsahují vyšší houby celiu řadu beta glukanů, které mají pozitivní vliv na lidksé zdraví. Projevuje se to zejména u 1,3 a 1,6 beta glukanů Stimulují leukocyty a zvyšují jejich množství v lidksé krvi. Snižují také množství cholesterolu a přispívají ke snižování tělesné hmotnosti. Účinné jsou např. pleuran z hlívy nebo lentinan z houževnatce., Together with chitin, the ?-glucans belong to building materials of mycetes cell walls. A high biological efficiency was found especially in ?-1,3- D-glucans and ?-1,6-D-glucans isolated from some basidiomycetes. These polysaccharides show a direct effect on leukocytes and stimulate them to produce antibodies and to increase their numbers in blood. Especially, they increase the numbers of Th1 lymphocytes, which have a positive effect on the protection of organism against allergic reactions. A number of ?-glucans, for example pleuran from Oyster (Pleurotus spp.) mushrooms or lentinan from Shi-Take (Lentinus edodes), showed a marked antioncogenic activity. In addition to an immunity-stimulating effect, ?-glucans may also participate in physiological processes related to the metabolism of fats in human body. Their application results in a decrease in the content of total cholesterol in blood and may also contribute to the reduction of body weight.

Microbial biofilms are a dominant feature of many human infections. However, developing effective strategies for controlling biofilms requires an understanding of the underlying biology well beyond what currently exists. Using a novel strategy, we have induced formation of a robust biofilm in Escherichia coli by utilizing an exogenous source of poly-N-acetylglucosamine (PNAG) polymer, a major virulence factor of many pathogens. Through microarray profiling of competitive selections, carried out in both transposon insertion and over-expression libraries, we have revealed the genetic basis of PNAG-based biofilm formation. Our observations reveal the dominance of electrostatic interactions between PNAG and surface structures such as lipopolysaccharides. We show that regulatory modulation of these surface structures has significant impact on biofilm formation behavior of the cell. Furthermore, the majority of clinical isolates which produced PNAG also showed the capacity to respond to the exogenously produced version of the polymer.

Barley is a desirable food ingredient, with health benefits provided by a p-glucan fibre fraction. A hull-less barley flour and flakes were incorporated into white and wholegrain wheat bread in quantities usually applied in practice. The breads were evaluated for nutritional composition and sensory properties and compared to standard products as controls. The supplemented breads were high in fibre, zinc and selenium content. It was estimated that a 300-gram daily portion of such breads could meet up to 40% of dietary recommended intakes for selenium and 70-75% of recommended daily values for beta-glucan. Regarding sensory quality, the only significant differences (p lt 0.05) were higher taste and lower volume in the white supplemented breads and lower crumb elasticity in the white bread made with barley flour. Hull-less barley can substantially contribute to an adequate intake of selenium and beta-glucan. In addition, supplemented breads were not found to pose a significant risk, with regard to excessive intakes of heavy elements (Pb, Cd, As).

Vedle chitinu obsahují vyšší houby celiu řadu beta glukanů, které mají pozitivní vliv na lidksé zdraví. Projevuje se to zejména u 1,3 a 1,6 beta glukanů Stimulují leukocyty a zvyšují jejich množství v lidksé krvi. Snižují také množství cholesterolu a přispívají ke snižování tělesné hmotnosti. Účinné jsou např. pleuran z hlívy nebo lentinan z houževnatce., Together with chitin, the ?-glucans belong to building materials of mycetes cell walls. A high biological efficiency was found especially in ?-1,3- D-glucans and ?-1,6-D-glucans isolated from some basidiomycetes. These polysaccharides show a direct effect on leukocytes and stimulate them to produce antibodies and to increase their numbers in blood. Especially, they increase the numbers of Th1 lymphocytes, which have a positive effect on the protection of organism against allergic reactions. A number of ?-glucans, for example pleuran from Oyster (Pleurotus spp.) mushrooms or lentinan from Shi-Take (Lentinus edodes), showed a marked antioncogenic activity. In addition to an immunity-stimulating effect, ?-glucans may also participate in physiological processes related to the metabolism of fats in human body. Their application results in a decrease in the content of total cholesterol in blood and may also contribute to the reduction of body weight.

The cholesterol-lowering effect of oats is well established, but the crucial properties eliciting this effect need to be further investigated to optimize the use of oats as functional foods. Furthermore, there are almost no reports investigating the effect of oats on atherosclerosis development. This thesis describes our work with finding suitable mouse models to study cholesterol-lowering and anti-atherogenic effects of oats, the mechanism behind, and how processing of oat foods might interfere with these beneficial effects. We found that supplementation of oat bran to an atherogenic diet significantly reduced plasma cholesterol and LDL+VLDL concentrations in C57BL/6 mice. The responsiveness to oats did however differ between two substrain of mice. Oat intake resulted in reduced plasma cholesterol, increased faecal excretion of bile acids and cholesterol, and increased expression of the bile acid producing enzyme CYP7A1 in the C57BL/6NCrl substrain. None of these parameters were altered in the C57BL/6JBomTac mice. The different expression of CYP7A1 in the two substrains of C57BL/6 strongly supports the importance of increased bile acid excretion, together with increase of bile acid synthesis from cholesterol, for oats to reduce levels of cholesterol in plasma. To address how processing of oats might interfere with its cholesterol-lowering properties, oat beta-glucans were enzymatically digested to different molecular weights and then fed to C57BL/6NCrl mice. Reducing the molecular weight of the beta-glucans affected its viscous properties in vitro. It also affected the production of short chain fatty acids in caecal contents of the mice, but did not influence the cholesterol-lowering properties. Thus molecular weight and viscous properties of beta-glucans do not seem to be crucial parameters for the cholesterol-lowering properties of oats in the C57BL/6 mice. When studying effects of oats on atherogenesis and inflammation we used a mouse model developing more prono

Autoimmunity is a complex trait disease where the environment influences susceptibility to disease by unclear mechanisms. T cell receptor clustering and signaling at the immune synapse, T cell proliferation, CTLA-4 endocytosis, T(H)1 differentiation, and autoimmunity are negatively regulated by beta1,6GlcNAc-branched N-glycans attached to cell surface glycoproteins. Beta1,6GlcNAc-branched N-glycan expression in T cells is dependent on metabolite supply to UDP-GlcNAc biosynthesis (hexosamine pathway) and in turn to Golgi N-acetylglucosaminyltransferases Mgat1, -2, -4, and -5. In Jurkat T cells, beta1,6GlcNAc-branching in N-glycans is stimulated by metabolites supplying the hexosamine pathway including glucose, GlcNAc, acetoacetate, glutamine, ammonia, or uridine but not by control metabolites mannosamine, galactose, mannose, succinate, or pyruvate. Hexosamine supplementation in vitro and in vivo also increases beta1,6GlcNAc-branched N-glycans in naïve mouse T cells and suppresses T cell receptor signaling, T cell proliferation, CTLA-4 endocytosis, T(H)1 differentiation, experimental autoimmune encephalomyelitis, and autoimmune diabetes in non-obese diabetic mice. Our results indicate that metabolite flux through the hexosamine and N-glycan pathways conditionally regulates autoimmunity by modulating multiple T cell functionalities downstream of beta1,6GlcNAc-branched N-glycans. This suggests metabolic therapy as a potential treatment for autoimmune disease.

Autoimmunity is a complex trait disease where the environment influences susceptibility to disease by unclear mechanisms. T cell receptor clustering and signaling at the immune synapse, T cell proliferation, CTLA-4 endocytosis, T(H)1 differentiation, and autoimmunity are negatively regulated by beta1,6GlcNAc-branched N-glycans attached to cell surface glycoproteins. Beta1,6GlcNAc-branched N-glycan expression in T cells is dependent on metabolite supply to UDP-GlcNAc biosynthesis (hexosamine pathway) and in turn to Golgi N-acetylglucosaminyltransferases Mgat1, -2, -4, and -5. In Jurkat T cells, beta1,6GlcNAc-branching in N-glycans is stimulated by metabolites supplying the hexosamine pathway including glucose, GlcNAc, acetoacetate, glutamine, ammonia, or uridine but not by control metabolites mannosamine, galactose, mannose, succinate, or pyruvate. Hexosamine supplementation in vitro and in vivo also increases beta1,6GlcNAc-branched N-glycans in naïve mouse T cells and suppresses T cell receptor signaling, T cell proliferation, CTLA-4 endocytosis, T(H)1 differentiation, experimental autoimmune encephalomyelitis, and autoimmune diabetes in non-obese diabetic mice. Our results indicate that metabolite flux through the hexosamine and N-glycan pathways conditionally regulates autoimmunity by modulating multiple T cell functionalities downstream of beta1,6GlcNAc-branched N-glycans. This suggests metabolic therapy as a potential treatment for autoimmune disease.

This thesis deals with the fermentation characteristics of lactic acid bacteria (LAB) in oat-based suspensions, with formulation work of fermented products based on oat and with nutritional studies of these products. Changes in structure in terms of viscosity and ropiness were studied when exopolysaccharide (EPS)-producing LAB strains, namely, Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772, Lactobacillus brevis G-77 and Pediococcus damnosus 2.6 were grown in these oat-based suspensions. Differences in structure were seen after growth of EPS-producing bacteria, resulting in higher viscosity and ropiness in these products. The formulation work of fermented products was carried out by using commercial yoghurt cultures with and without the presence of an EPS-producing strain. The products developed were found to have sensory acceptance and good bacterial survival during storage. The bacterial survival of three probiotic strains, Lactobacillus reuteri ATCC 55730, Lactobacillus acidophilus DSM 20079 and Bifidobacterium bifidum DSM 20456, was also studied in different oat-based suspensions with and without the influence of a yoghurt culture during 30 days of storage. The highest survival was seen for the Lactobacillus reuteri strain. The co-fermentation in the presence of a yoghurt culture gave a lower pH in the final products and decreased the survival rates for all strains. The nutritional effects of fermented, oat-based products were investigated by using germfree and conventional rats and by performing a clinical study on 56 healthy subjects. Physiological parameters such as changes in serum cholesterol, faecal excretion of cholesterol, and amount of faecal short chain fatty acids (SCFA) were studied. In addition, changes in the faecal flora were investigated in the human subjects. No changes in the serum cholesterol levels or faecal excretion of cholesterol were seen in the rat models when the animals were fed on different, fermented oat-based products. A diffe

β1,6-Glucan is a key component of the yeast cell wall, interconnecting cell wall proteins, β1,3-glucan, and chitin. It has been postulated that the synthesis of β1,6-glucan begins in the endoplasmic reticulum with the formation of protein-bound primer structures and that these primer structures are extended in the Golgi complex by two putative glucosyltransferases that are functionally redundant, Kre6 and Skn1. This is followed by maturation steps at the cell surface and by coupling to other cell wall macromolecules. We have reinvestigated the role of Kre6 and Skn1 in the biogenesis of β1,6- glucan. Using hydrophobic cluster analysis, we found that Kre6 and Skn1 show significant similarities to family 16 glycoside hydrolases but not to nucleotide diphospho-sugar glycosyltransferases, indicating that they are glucosyl hydrolases or transglucosylases instead of genuine glucosyltransferases. Next, using immunogold labeling, we tried to visualize intracellular β1,6-glucan in cryofixed sec1-1 cells which had accumulated secretory vesicles at the restrictive temperature. No intracellular labeling was observed, but the cell surface was heavily labeled. Consistent with this, we could detect substantial amounts of β1,6-glucan in isolated plasma membrane-derived microsomes but not in post-Golgi secretory vesicles. Taken together, our data indicate that the synthesis of β1,6-glucan takes place largely at the cell surface. An alternative function for Kre6 and Skn1 is discussed.

The GGP1/GAS1 gene codes for a glycosylphosphatidylinositol-anchored plasma membrane glycoprotein of Saccharomyces cerevisiae. The ggp1delta mutant shows morphogenetic defects which suggest changes in the cell wall matrix. In this work, we have investigated cell wall glucan levels and the increase of chitin in ggp1delta mutant cells. In these cells, the level of alkali-insoluble 1,6-beta-D-glucan was found to be 50% of that of wild-type cells and was responsible for the observed decrease in the total alkali-insoluble glucan. Moreover, the ratio of alkali-soluble to alkali-insoluble glucan almost doubled, suggesting a change in glucan solubility. The increase of chitin in ggp1delta cells was found to be essential since the chs3delta ggp1delta mutations determined a severe reduction in the growth rate and in cell viability. Electron microscopy analysis showed the loss of the typical structure of yeast cell walls. Furthermore, in the chs3delta ggp1delta cells, the level of alkali-insoluble glucan was 57% of that of wild-type cells and the alkali-soluble/alkali-insoluble glucan ratio was doubled. We tested the effect of inhibition of chitin synthesis also by a different approach. The ggp1delta cells were treated with nikkomycin Z, a well-known inhibitor of chitin synthesis, and showed a hypersensitivity to this drug. In addition, studies of genetic interactions with genes related to the construction of the cell wall indicate a synthetic lethal effect of the ggp1delta kre6delta and the ggp1delta pkc1delta combined mutations. Our data point to an involvement of the GGP1 gene product in the cross-links between cell wall glucans (1,3-beta-D-glucans with 1,6-beta-D-glucans and with chitin). Chitin is essential to compensate for the defects due to the lack of Ggp1p. Moreover, the activities of Ggp1p and Chs3p are essential to the formation of the organized structure of the cell wall in vegetative cells

Bakalářská práce se zabývá charakterizací přírodních beta-glukanů a dalších aktivních látek izolovaných z makrořas a kvasinek. Použité vzorky makrořas se od sebe liší pigmentem, původem a metodou extrakce. Teoretická část se zaměřuje na druhy mořských řas a kvasinek, na beta-glukany, jejich zdroje a biologické účinky, dále na další biologicky aktivní látky (jako jsou např.: polyfenoly a antioxidanty) a nakonec na metody stanovení těchto biologicky aktivních látek. V praktické časti byly analyzovány vzorky vybraných makrořas, konkrétně vzorky Arame, Nori, Kombu, Wakame, Hijiki a Spaghetti. V práci byly připraveny vodné, ethanolové a acetonové extrakty těchto řas. Vodné extrakty byly použity pro spektrofotometrické stanovení fenolyckých látek, flavonoidů a antioxidační aktivity metodou ABTS. Ethanolové a acetonové extrakty zmíněných makrořas byly použity pro spektrofotometrické stanovení obsahu celkového chlorofylu a karotenoidů. Součásti práce byla také analýza obsahu beta-glukanů u makrořas a kvasinky R. toruloides. Vysoký obsah beta-glukanů měly řasy Arame a Nori a kvasinky R. toruloides. Obsah beta-glukanů byl v případě ostatních makrořas velice nízký., The bachelor thesis deals with the characterization of natural beta-glucans and other active substances isolated from macroalgae and yeasts. The used macroalgae samples differ in the pigment, origin and method of algae processing. The theoretical part focuses on seaweed and yeast species, beta-glucans, their sources and biological effects, other biologically active substances (such as polyphenols and antioxidants) and finally on methods for determining these biologically active substances. In the practical part, samples of selected macroalgae were analyzed, namely samples of Arame, Nori, Kombu, Wakame, Hijiki and Spaghetti. Aqueous, ethanol and acetone extracts of these algae were prepared in this work. The aqueous extracts were used for spectrophotometric determination of phenolic substances, flavonoids and antioxidant activity by the ABTS method. Furthermore, ethanol and acetone extracts of said macroalgae were used to determine total chlorophyll and carotenoid content spectrophotometric. The work also focused on beta-glucan content, analyzing macroalgae and yeast R. toruloides. Arame and Nori algae and R. toruloides were high in beta-glucans. However, beta-glucans were very low in other macroalgae.

Tato bakalářská práce je zaměřena na studium vlivu různých kultivačních podmínek na produkci biomasy, beta-glukanů, lipidů, karotenoidů a dalších metabolitů u vybraných kmenů karotenogenních kvasinek. Práce je rozdělena na dvě hlavní části, a to část teoretickou a část experimentální. V teoretické části jsou charakterizovány beta-glukany, možnosti jejich zdrojů a využití. Dále jsou zde popsány zvolené rody kvasinek, další jimi produkované metabolity, a nakonec možné metody analýzy těchto látek. V experimentální části je popsán postup kultivace, analýzy a získané výsledky. V rámci prvního experimentu byly zvolené kmeny kvasinek kultivovány na produkčních médiích s různým C/N poměrem, během druhého experimentu byly kvasinky kultivovány při různém obsahu fosforu, a to buď za laboratorní nebo za snížené teploty. Zvoleny byly kmeny Rhodotorula glutinis, Rhodotorula mucilaginosa, Rhodosporidium toruloides, Cystofilobasidium macerans, Sporidiobolus metaroseus a Sporidiobolus pararoseus. Zvyšující se C/N poměr měl obecně pozitivní vliv na produkci lipidů, zato u karotenoidů působil opačným způsobem, na množství beta-glukanů neměl jednoznačný vliv. Bylo zjištěno, že snížení kultivační teploty má výrazně negativní efekt na produkci všech sledovaných metabolitů. Zvýšený obsah fosforu v produkčních médiích u většiny kmenů pozitivně ovlivnil produkci biomasy a zastoupení beta-glukanů, naopak největších hodnot obsahu karotenoidů bylo dosaženo při nižší koncentraci fosforu. Na množství a zastoupení lipidů neměl obsah fosforu téměř žádný vliv. Nejvyšší zastoupení beta-glukanů v biomase bylo zjištěno u kmene C. macerans, jejich největší produkce dosáhl kmen R. toruloides. Pro akumulaci lipidů v biomase byl nejvhodnější kmen S. metaroseus. Nejvyšší množství celkových karotenoidů bylo dosaženo u kmene R. mucilaginosa., The aim of this bachelor thesis is to study the effect of different cultivation conditions on the production of biomass, beta-glucans, lipids, carotenoids and other metabolites in selected strains of carotenogenic yeast. The thesis is divided into two main parts, a theoretical one and an experimental one. Theoretical part contains a characterisation of beta-glucans, their sources and uses, furthermore a description of the selected yeast genera and other metabolites they produce and finally possible analytical methods of these compounds. Experimental part describes the cultivation, analysis and obtained results. During the first experiment, the yeast strains were cultivated in production media with different C/N ratio, during the second experiment, the yeasts were cultivated either at room or at lower temperature with different content of phosphorus in the media. The selected strains of yeast were Rhodotorula glutinis, Rhodotorula mucilaginosa, Rhodosporidium toruloides, Cystofilobasidium macerans, Sporidiobolus metaroseus and Sporidiobolus pararoseus. Increasing C/N ratio generally had a positive effect on the production of lipids, while exhibiting the opposite effect on carotenoids, it did not have an unequivocal effect on the amount of beta-glucans. It was discovered that lowering the cultivation temperature had a significant negative effect on the production of all observed metabolites. Increased content of phosphorus in production media positively influenced the production of biomass and beta-glucans in most strains, on the other hand the highest amount of carotenoids was reached at lower concentration of phosphorus. The amount of phosphorus had almost no effect on the production and the profile of lipids. The highest amount of beta-glucans in biomass was discovered in C. macerans, their biggest production was reached by R. toruloides. The strain S. metaroseus was the most suitable for accumulation of lipids in its biomass. The highest amount of total carotenoid

Bakalářská práce se zabývá charakterizací přírodních beta-glukanů a dalších aktivních látek izolovaných z makrořas a kvasinek. Použité vzorky makrořas se od sebe liší pigmentem, původem a metodou extrakce. Teoretická část se zaměřuje na druhy mořských řas a kvasinek, na beta-glukany, jejich zdroje a biologické účinky, dále na další biologicky aktivní látky (jako jsou např.: polyfenoly a antioxidanty) a nakonec na metody stanovení těchto biologicky aktivních látek. V praktické časti byly analyzovány vzorky vybraných makrořas, konkrétně vzorky Arame, Nori, Kombu, Wakame, Hijiki a Spaghetti. V práci byly připraveny vodné, ethanolové a acetonové extrakty těchto řas. Vodné extrakty byly použity pro spektrofotometrické stanovení fenolyckých látek, flavonoidů a antioxidační aktivity metodou ABTS. Ethanolové a acetonové extrakty zmíněných makrořas byly použity pro spektrofotometrické stanovení obsahu celkového chlorofylu a karotenoidů. Součásti práce byla také analýza obsahu beta-glukanů u makrořas a kvasinky R. toruloides. Vysoký obsah beta-glukanů měly řasy Arame a Nori a kvasinky R. toruloides. Obsah beta-glukanů byl v případě ostatních makrořas velice nízký., The bachelor thesis deals with the characterization of natural beta-glucans and other active substances isolated from macroalgae and yeasts. The used macroalgae samples differ in the pigment, origin and method of algae processing. The theoretical part focuses on seaweed and yeast species, beta-glucans, their sources and biological effects, other biologically active substances (such as polyphenols and antioxidants) and finally on methods for determining these biologically active substances. In the practical part, samples of selected macroalgae were analyzed, namely samples of Arame, Nori, Kombu, Wakame, Hijiki and Spaghetti. Aqueous, ethanol and acetone extracts of these algae were prepared in this work. The aqueous extracts were used for spectrophotometric determination of phenolic substances, flavonoids and antioxidant activity by the ABTS method. Furthermore, ethanol and acetone extracts of said macroalgae were used to determine total chlorophyll and carotenoid content spectrophotometric. The work also focused on beta-glucan content, analyzing macroalgae and yeast R. toruloides. Arame and Nori algae and R. toruloides were high in beta-glucans. However, beta-glucans were very low in other macroalgae.

Tato bakalářská práce je zaměřena na studium vlivu různých kultivačních podmínek na produkci biomasy, beta-glukanů, lipidů, karotenoidů a dalších metabolitů u vybraných kmenů karotenogenních kvasinek. Práce je rozdělena na dvě hlavní části, a to část teoretickou a část experimentální. V teoretické části jsou charakterizovány beta-glukany, možnosti jejich zdrojů a využití. Dále jsou zde popsány zvolené rody kvasinek, další jimi produkované metabolity, a nakonec možné metody analýzy těchto látek. V experimentální části je popsán postup kultivace, analýzy a získané výsledky. V rámci prvního experimentu byly zvolené kmeny kvasinek kultivovány na produkčních médiích s různým C/N poměrem, během druhého experimentu byly kvasinky kultivovány při různém obsahu fosforu, a to buď za laboratorní nebo za snížené teploty. Zvoleny byly kmeny Rhodotorula glutinis, Rhodotorula mucilaginosa, Rhodosporidium toruloides, Cystofilobasidium macerans, Sporidiobolus metaroseus a Sporidiobolus pararoseus. Zvyšující se C/N poměr měl obecně pozitivní vliv na produkci lipidů, zato u karotenoidů působil opačným způsobem, na množství beta-glukanů neměl jednoznačný vliv. Bylo zjištěno, že snížení kultivační teploty má výrazně negativní efekt na produkci všech sledovaných metabolitů. Zvýšený obsah fosforu v produkčních médiích u většiny kmenů pozitivně ovlivnil produkci biomasy a zastoupení beta-glukanů, naopak největších hodnot obsahu karotenoidů bylo dosaženo při nižší koncentraci fosforu. Na množství a zastoupení lipidů neměl obsah fosforu téměř žádný vliv. Nejvyšší zastoupení beta-glukanů v biomase bylo zjištěno u kmene C. macerans, jejich největší produkce dosáhl kmen R. toruloides. Pro akumulaci lipidů v biomase byl nejvhodnější kmen S. metaroseus. Nejvyšší množství celkových karotenoidů bylo dosaženo u kmene R. mucilaginosa., The aim of this bachelor thesis is to study the effect of different cultivation conditions on the production of biomass, beta-glucans, lipids, carotenoids and other metabolites in selected strains of carotenogenic yeast. The thesis is divided into two main parts, a theoretical one and an experimental one. Theoretical part contains a characterisation of beta-glucans, their sources and uses, furthermore a description of the selected yeast genera and other metabolites they produce and finally possible analytical methods of these compounds. Experimental part describes the cultivation, analysis and obtained results. During the first experiment, the yeast strains were cultivated in production media with different C/N ratio, during the second experiment, the yeasts were cultivated either at room or at lower temperature with different content of phosphorus in the media. The selected strains of yeast were Rhodotorula glutinis, Rhodotorula mucilaginosa, Rhodosporidium toruloides, Cystofilobasidium macerans, Sporidiobolus metaroseus and Sporidiobolus pararoseus. Increasing C/N ratio generally had a positive effect on the production of lipids, while exhibiting the opposite effect on carotenoids, it did not have an unequivocal effect on the amount of beta-glucans. It was discovered that lowering the cultivation temperature had a significant negative effect on the production of all observed metabolites. Increased content of phosphorus in production media positively influenced the production of biomass and beta-glucans in most strains, on the other hand the highest amount of carotenoids was reached at lower concentration of phosphorus. The amount of phosphorus had almost no effect on the production and the profile of lipids. The highest amount of beta-glucans in biomass was discovered in C. macerans, their biggest production was reached by R. toruloides. The strain S. metaroseus was the most suitable for accumulation of lipids in its biomass. The highest amount of total carotenoid