Non-conventional wine yeasts are extensively studied as promising producers of hydrolytic enzymes and as potential starter cultures in winemaking due to their ability to improve organoleptic properties of wine. Thirty-six yeast strains of enological and brewery origin from the Ukrainian Collection of Microorganisms belonging to Torulaspora, Kloeckera, Candida, Metschnikowia, Pichia, and Zygosaccharomyces genera have been screened for the production of extracellular hydrolases, stress tolerance, fermentative activity, and other traits of enological interest. This study revealed the high incidence of lipolytic, proteolytic, and β-glucosidase activities among the yeasts, while no pectinase activity was detected. Esterase, cellulase and glucanase activities were found in a small proportion of yeasts (8.33-16.66%). Several Pichia anomala, Kloekera javanica, Pichia membranifaciens, and Metschnikowia pulcherrima strains demonstrated a wide range of hydrolytic activities. High tolerance to stress factors (ethanol, osmotic, and oxidative stress) present during alcoholic fermentation was detected in P. anomala and M. pulcherrima strains. Fermentative activity of several yeast strains was evaluated in microfermentations in a model semi-synthetic medium. Strain P. anomala UCM Y-216 was selected as the most promising culture for winemaking due to its hydrolytic activities, tolerance to stress factors and other valuable metabolic traits. This study represents the first step for selecting a non-conventional yeast strain of enological origin as a potential co-culture for winemaking.
The red yeasts are currently widely discussed and controversial group of yeasts because of the growing number of reports of their ability to become opportunistic pathogens of plants, animals and humans. The aim of this work was complex identification of the red yeast culture isolated from gastrointestinal tract of healthy Hucul long-liver from the Carpathians highland region of Ukraine. Torularhodin was found to be a major component within yeast culture carotenoids complex. According to conventional biochemical and morphological approaches as well as to molecular biological investigation of internal transcribed spacer region (ITS) of ribosomal operon it was concluded that isolate belonged to species Rhodotorula mucilaginosa.
Hydrolysis of lignocellulose to fermentable sugars and their subsequent conversion to ethanol remain great challenges in the biofuel industry. Rotten wood is first colonized by bacteria and molds that possess strong hydrolases. Yeasts are also an important group of microorganisms that may participate in wood hydrolysis. Decaying wood could provide a rich natural reservoir of yeasts possessing promising hydrolytic activities, including xylanases, cellulases, β-glucosidases, or abilities essential for the fermentation of pentose sugars derived from lignocellulose degradation, especially xylose. Therefore, the aim of this work was to screen yeasts isolated from rotten wood samples for the production of hydrolytic enzymes directed at lignocellulose components and the ability to ferment xylose, L-arabinose, and cellobiose. Methods. Yeast strains were isolated from 22 samples of rotten wood and identified by phenotypic characteristics according to Kurtzman et al. Hydrolytic properties and the ability of the isolated strains to ferment xylose, L-arabinose, and cellobiose were determined using conventional methods. Results. 30 strains of yeasts and yeast-like micromycetes were isolated from 22 samples of rotten wood in the Holosiivskyi Forest, Kyiv. Based on phenotypic properties, most of the isolated yeasts belonged to ascomycetous yeasts and were represented by the following genera: Candida (8 strains), Debaryomyces (5 strains), Kluyveromyces (5 strains), Pichia (5 strains), Scheffersomyces (2 strains), Lachancea, Hanseniaspora, Saccharomyces, and Geotrichum/Galactomyces. A strain of yeast-like non-photosynthetic alga Prototheca sp. was also detected. Most of the isolated microfungi (66.6% isolates) exhibited extracellular β-glucosidase activity, two Candida tropicalis strains possessed weak pectinase and xylanase activity. None of the isolates demonstrated extracellular cellulase activity. Two yeast strains preliminarily identified as Scheffersomyces stipitis were able to ferment xylose at a concentration of 20—100 g/L over a wide temperature range up to 37°C. Acetic acid at 0.25—1% (v/v) concentration resulted in the complete inhibition of xylose fermentation. Ethanol production from xylose up to 6 g/L was observed under the microaerobic fermentation conditions for 24 hr at the substrate concentration 40 g/L, but the subsequent fermentation resulted in decreasing ethanol concentration presumably due to ethanol re-assimilation. None of the isolated strains was capable of fermenting cellobiose or L-arabinose under the microaerobic conditions. Conclusions. This work provides the characterization of yeast microbiota of rotten wood that was represented predominantly by ascomycetous yeasts. The dominant extracellular hydrolytic activity of the isolates was β-glucosidase. This is the first report on the isolation of xylose-fermenting yeasts Scheffersomyces stipitis in Ukraine, which comprised 7% of all the microfungi isolated from rotten wood.
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