Beta (β)-glucans are polysaccharides composed of D-glucose monomers. Nowadays, β-glucans are gaining attention due to their attractive immunomodulatory biological activities, which can be utilized in pharmaceutical or food supplementation industries. Some carotenogenic Basidiomycetes yeasts, previously explored for lipid and carotenoid coproduction, could potentially coproduce a significant amount of β-glucans. In the present study, we screened eleven Basidiomycetes for the coproduction of lipids and β-glucans. We examined the effect of four different C/N ratios and eight different osmolarity conditions on the coproduction of lipids and β-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 strains C. infirmominiatum CCY 17-18-4 and R. kratochvilovae CCY 20-2-26 were identified as the best coproducers of lipids and β-glucans. In addition, C. infirmominiatum CCY 17-18-4, R. kratochvilovae CCY 20-2-26 and P. rhodozyma CCY 77-1-1 were identified as the best alternative producers of β-glucans. Increased C/N ratio led to increased biomass, lipid and β-glucans production for several yeast strains. Increased osmolarity had a negative effect on biomass and lipid production while the β-glucan production was positively affected.

• Klíčová slova
• carbon:nitrogen ratio, high-throughput screening, lipids, osmotic stress, red yeast, β-glucans,
• Publikační typ
• časopisecké články MeSH

Carotenogenic yeasts are non-conventional oleaginous microorganisms capable of utilizing various waste substrates. In this work, four red yeast strains (Rhodotorula, Cystofilobasidium, and Sporobolomyces sp.) were cultivated in media containing crude, emulsified, and enzymatically hydrolyzed animal waste fat, compared with glucose and glycerol, as single C-sources. Cell morphology (cryo-SEM (cryo-scanning electron microscopy), TEM (transmission electron microscopy)), production of biomass, lipase, biosurfactants, lipids (gas chromatography/flame ionization detection, GC/FID) carotenoids, ubiquinone, and ergosterol (high performance liquid chromatography, HPLC/PDA) in yeast cells was studied depending on the medium composition, the C source, and the carbon/nitrogen (C/N) ratio. All studied strains are able to utilize solid and processed fat. Biomass production at C/N = 13 was higher on emulsified/hydrolyzed fat than on glucose/glycerol. The production of lipids and lipidic metabolites was enhanced for several times on fat; the highest yields of carotenoids (24.8 mg/L) and lipids (54.5%/CDW (cell dry weight)) were found in S. pararoseus. Simultaneous induction of lipase and biosurfactants was observed on crude fat substrate. An increased C/N ratio (13-100) led to higher biomass production in fat media. The production of total lipids increased in all strains to C/N = 50. Oppositely, the production of carotenoids, ubiquinone, and ergosterol dramatically decreased with increased C/N in all strains. Compounds accumulated in stressed red yeasts have a great application potential and can be produced efficiently during the valorization of animal waste fat under the biorefinery concept.

• Klíčová slova
• biosurfactants, carotenogenic yeasts, carotenoids, ergosterol, lipase, lipids, ubiquinone,
• Publikační typ
• časopisecké články MeSH

Cardiolipin and phosphatidylglycerol are anionic phospholipids localized to the inner mitochondrial membrane. In this study, it is demonstrated by fluorescence and transmission electron microscopy that atp2.1pgs1Δ mutant mitochondria lacking anionic phospholipids contain fragmented and swollen mitochondria with a completely disorganized inner membrane. In the second part of this study, it was shown that the temperature sensitivity of the atp2.1pgs1Δ mutant was not suppressed by the osmotic stabilizer glucitol but by glucosamine, a precursor of chitin synthesis. The atp2.1pgs1Δ mutant was hypersensitive to Calcofluor White and caffeine, resistant to Zymolyase, but its sensitivity to caspofungin was the same as the strains with the standard PGS1 gene. The distribution of chitin in the mutant cell wall was impaired. The glucan level in the cell wall of the atp2.1pgs1Δ mutant was reduced by 4-8 %, but the level of chitin was almost double that in the wild-type strain. The cell wall of the atp2.1pgs1Δ mutant was about 20 % thinner than the wild type, but its morphology was not significantly altered.

• MeSH
• aerobióza MeSH
• buněčná stěna chemie   ultrastruktura   MeSH
• delece genu MeSH
• fluorescenční mikroskopie MeSH
• fosfolipidy nedostatek   MeSH
• glukany analýza   MeSH
• Kluyveromyces cytologie   genetika   růst a vývoj   metabolismus   MeSH
• mitochondrie ultrastruktura   MeSH
• teplota MeSH
• transmisní elektronová mikroskopie MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fosfolipidy MeSH
• glukany MeSH

The SUN family is comprised of proteins that are conserved among various yeasts and fungi, but that are absent in mammals and plants. Although the function(s) of these proteins are mostly unknown, they have been linked to various, often unrelated cellular processes such as those connected to mitochondrial and cell wall functions. Here we show that three of the four Saccharomyces cerevisiae SUN family proteins, Uth1p, Sim1p and Sun4p, are efficiently secreted out of the cells in different growth phases and their production is affected by the level of oxygen. The Uth1p, Sim1p, Sun4p and Nca3p are mostly synthesized during the growth phase of both yeast liquid cultures and colonies. Culture transition to slow-growing or stationary phases is linked with a decreased cellular concentration of Sim1p and Sun4p and with their efficient release from the cells. In contrast, Uth1p is released mainly from growing cells. The synthesis of Uth1p and Sim1p, but not of Sun4p, is repressed by anoxia. All four proteins confer cell sensitivity to zymolyase. In addition, Uth1p affects cell sensitivity to compounds influencing cell wall composition and integrity (such as Calcofluor white and Congo red) differently when growing on fermentative versus respiratory carbon sources. In contrast, Uth1p is essential for cell resistance to boric acids irrespective of carbon source. In summary, our novel findings support the hypothesis that SUN family proteins are involved in the remodeling of the yeast cell wall during the various phases of yeast culture development and under various environmental conditions. The finding that Uth1p is involved in cell sensitivity to boric acid, i.e. to a compound that is commonly used as an important antifungal in mycoses, opens up new possibilities of investigating the mechanisms of boric acid's action.

• MeSH
• benzensulfonáty metabolismus   farmakologie   MeSH
• buněčná stěna metabolismus   MeSH
• extracelulární prostor metabolismus   MeSH
• glukosidasy genetika   metabolismus   MeSH
• intracelulární prostor metabolismus   MeSH
• kyseliny borité metabolismus   farmakologie   MeSH
• membránové proteiny genetika   metabolismus   MeSH
• mitochondriální proteiny genetika   metabolismus   MeSH
• proteiny teplotního šoku genetika   metabolismus   MeSH
• regulace genové exprese u hub MeSH
• represorové proteiny genetika   metabolismus   MeSH
• Saccharomyces cerevisiae - proteiny genetika   metabolismus   MeSH
• Saccharomyces cerevisiae účinky léků   genetika   růst a vývoj   metabolismus   MeSH
• spotřeba kyslíku * MeSH
• transkripční faktory bHLH genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• benzensulfonáty MeSH
• C.I. Fluorescent Brightening Agent 28 MeSH Prohlížeč
• glukosidasy MeSH
• kyseliny borité MeSH
• membránové proteiny MeSH
• mitochondriální proteiny MeSH
• proteiny teplotního šoku MeSH
• represorové proteiny MeSH
• Saccharomyces cerevisiae - proteiny MeSH
• SIM1 protein, S cerevisiae MeSH Prohlížeč
• Sun4 protein, S cerevisiae MeSH Prohlížeč
• transkripční faktory bHLH MeSH
• UTH1 protein, S cerevisiae MeSH Prohlížeč