Xylan-degrading activity in yeasts: Growth on xylose,xylan and hemicelluloses

The ability to grow in liquid media withD-xylose, xylan from deciduous trees, and hemicelluloses from conifers was tested in 95 strains of 35 genera of yeasts and yeast-like organisms. Of 54 strains thriving on xylose, only 13 (generaAureobasidium, Cryptococcus andTrichosporon) utilized xylan and hemicelluloses as growth substrates. The árowth media of these strains were found to contain xylandegrading enzymes splitting the substrate to xylose and a mixture of xylose oligosaccharides. The ability of these yeasts to utilize the wood components (hitherto unknown in the genusCryptococeus) makes them potential producers of microbial proteins from industrial wood wastes containing xylose oligosaccharides, xylan, and hemicelluloses as the major saccharide components without previous saccharification.     

The application of whole-cell protein electrophoresis for the classification and identification of basidiomycetous yeast species

The relationships among 65 basidiomycetous yeast strains were determined by one-dimensional electrophoresis of SDS-solubilized whole-cell proteins. Protein profiles were compared by the Pearson product moment correlation coefficient (r). The strains investigated represented species from the generaCystofilobasidium, Filobasidium, Filobasidiella, Kondoa, Leucosporidium, Mrakia andRhodosporidium. Except for the genusMrakia, all species constituted separate protein electrophoretic clusters. The species of the genusMrakia (M. frigida, M. gelida, M. nivalis andM. stokesii) show highly similar protein patterns, suggesting that these four species may be synonymous. Strains of two varieties ofFilobasidiella neoformans, F. neoformans var.neoformans andF. neoformans var.bacillispora, could not be differentiated by protein electrophoresis.For the delineation of the protein electrophoretic clusters of the yeasts studied, literature data relying on other criteria, such as DNA base composition, carbon source utilization patterns, enzymatic protein electrophoregrams, ubiquinone systems, DNA-DNA homology and rRNA sequence data were used. It was demonstrated that a database of SDS-protein patterns provides a valuable tool for the identification of yeasts.     

Degradation of Nucleic Acids and their Related Compounds by Microbial Enzymes

Attempts were made to form 5′-mononucleotides from non-proliferating cells of various kinds of yeasts. It was found that yeasts were devided into four groups according to their ability to excrete compounds absorbing at 260 mμ, the first of which excreted UV-absorbing materials at the acid pH (I), the second at the alkaline pH (II), the third both at the acid and alkaline pHs (III), and the fourth showed weak ability to excrete UV-absorbing materials at both pHs (IV). In general, 3′-mononucleotides were excreted at the acid pH and 5′-mononucleotides at the alkaline pH. Rhodotorula pallida, however, excreted 5′-mononucleotides both at the acid and alkaline pHs.     

Assimilation of benzoate byRhodotorula rubra,Rhodotorula glutinis andRhodosporidium toruloides,as affected by glucose or xylose

Xylose or glucose (5 g/l) was utilized simultaneously with benzoate (5 g/l) byRhodosporidium toruloides andRhodotorula rubra in batch culture. At a higher glucose concentration, benzoate was utilized only after glucose was depleted from the media. Both yeasts preferentially utilized benzoate before xylose even if there were more than 5 g xylose/l.Rhodotorula glutinis preferentially utilized glucose (10 g/l) before benzoate but utilized xylose and benzoate simultaneously.The authors are with the Department of Biochemical Technology, Faculty of Chemistry, Slovak Technical University, Radlinského 9, 812 37 Bratislava, Slovak Republic     

Extracytoplasmic phosphatases of selected species of Saccharomyces, Kluyveromyces and Rhodotorula

Phosphatase activities of yeasts belonging to the genera Saccharomyces, Kluyveromyces and Rhodotorula were studied. Rhodotorula rubra exhibited activities at acid, neutral and alkaline pH; the other yeasts only had activity at acid pH. Growing yeasts in a constant pH (4.5) medium decreased phosphatase activities in Saccharomyces and Kluyveromyces, while neutral activity was enhanced in Rhodotorula rubra which excreted more enzyme under these conditions. Washing cells with sucrose solutions lowered phosphatase activities in all yeasts, due to enzyme liberation. Acid phosphatase activities in isolated and purified cell walls were very small. Phosphatases thus appear not to be strongly bound to yeast cell walls.     

云南高原湖泊杞麓湖冬季可培养酵母菌多样性分析

【目的】探究云南杞麓湖酵母菌群落结构及其与环境因子的相互关系。【方法】采用原位培养方法对杞麓湖14个水样进行酵母菌分离,应用26S r DNA D1/D2区域序列分析,并结合形态及生理生化指标将对分离获得的酵母菌进行鉴定,运用软件bio-dap和Canoco分析酵母菌类群的丰富度及其与环境因子间的相互关系。【结果】从杞麓湖中分离得到321株酵母菌,鉴定为14个属27个种和1个潜在的新类群。Rhodosporidium kratochvilovae和出芽短梗霉(Aureobasidium pullulans)是优势种,分别为总菌株数的29.6%和16.8%。水体总磷含量是影响产色素红冬孢酵母属(Rhodosporidium)分布的重要环境因子,而p H为隐球酵母属(Cryptococcus)分布的一个重要选择条件。【结论】杞麓湖酵母菌具有较为丰富的群落多样性。     

Yeast contaminants of micropropagated plant cultures

L eifert , C., W aites , W.M., N icholas , J.R. & K eetley , J.W. 1990. Yeast contaminants of micropropagated plant cultures. Journal of Applied Bacteriology 69 , 471–476.
Of 36 yeast strains isolated from contaminated plant cultures 78% were Candida, 20% Rhodotorula and 2% were Cryptococcus species. Strains of Candida guilliermon-dii represented 45% of all yeasts isolated. Yeasts grew rapidly on plant growth media with a pH of between 2.5 and 6.0 and decreased the medium pH to values of between 2.0 and 3.0. Candida yeasts growing on plant medium for 28 d metabolized about 75% of the sucrose and produced fermentation products such as ethanol and acetic acid. In contrast, Rhodotorula did not produce ethanol or acetic acid and only metabolized about 20% of the sucrose. Possible sources of contamination are discussed.     

Studies on the effect of hydrogen-ion concentration on the growth and sporulation of certain soil fungi

Summary A comparative study on the effect of pH, on growth and sporulation of the strains ofAspergillus terreus andCurvularia lunata obtained from alkaline and fertile soils andHeterosporium allii from alkaline soils has been made. The optimum and maximum pH for growth and sporulation has been found to be higher for the forms isolated from alkaline soils.Portion of a thesis presented at the University of Lucknow for the degree of Doctor of Philosophy.     

Oily yeasts as oleaginous cell factories

Oily yeasts have been described to be able to accumulate lipids up to 20% of their cellular dry weight. These yeasts represent a minor proportion of the total yeast population, and only 5% of them have been reported as able to accumulate more than 25% of lipids. The oily yeast genera include Yarrowia, Candida, Rhodotorula, Rhodosporidium, Cryptococcus, Trichosporon, and Lipomyces. More specifically, examples of oleaginous yeasts include the species: Lipomyces starkeyi, Rhodosporidium toruloides, Rhodotorula glutinis, and Yarrowia lipolytica. Yeast do exhibit advantages for lipid production over other microbial sources, namely, their duplication times are usually lower than 1 h, are much less affected than plants by season or climate conditions, and their cultures are more easily scaled up than those of microalgae. Additionally, some oily yeasts have been reported to accumulate oil up to 80% of their dry weight and can indeed generate different lipids from different carbon sources or from lipids present in the culture media. Thus, they can vary their lipid composition by replacing the fatty acids present in their triglycerides. Due to the diversity of microorganisms and growth conditions, oily yeasts can be useful for the production of triglycerides, surfactants, or polyunsaturated fatty acids.     

Movements of protons coupled to glucose transport in yeasts. A comparative study among 248 yeast strains

In 248 strains representing 205 yeast species, changes in pH coupled to glucose addition were followed in unbuffered cell suspensions. Alkalinization of the external medium elicited by glucose, indicating a H⁺-glucose symport was observed in 34% of the strains, most of them belonging to the genera Rhodotorula, Hansenula and Candida. H⁺ uptake coupled to glucose transport was observed only after exhaustion of glucose in growth media. This observation was taken as an indication that, in general, the synthesis of H⁺-glucose symport is under the control of catabolite repression. Subsequently to the addition of glucose, in most yeasts (82%) acidification was observed. This ability is probably related to the creation of a proton-gradient across the plasma membrane and is generally distributed among yeasts.     

Production of extracellular polymers by yeast-like generaDipodascus andDipodascopsis under NaCl stress

Changes in the production and basal characteristics of extracellular polymeric compounds were analyzed in eight yeast-like fungi of the generaDipodascus andDipodascopsis in media with and without the addition of NaCl. In the presence of higher concentration of NaCl (8%) the strains produced extracellular polymers (EP) of relatively lower molar mass. Stress EP contained a higher percentage of glutamic acid, mannose and galactose than normal EP. The production of these compounds is probably part of the adaptation mechanism and extracellular protection against disbalanced growth.     

Spores of the mycorrhizal fungus <Emphasis Type="Italic">Glomus mosseae</Emphasis> host yeasts that solubilize phosphate and accumulate polyphosphates

The present paper reports the presence of bacteria and yeasts tightly associated with spores of an isolate of Glomus mosseae. Healthy spores were surface disinfected by combining chloramine-T 5%, Tween-40, and cephalexin 2.5 g L⁻¹ (CTCf). Macerates of these spores were incubated on agar media, microorganisms were isolated, and two yeasts were characterized (EndoGm1, EndoGm11). Both yeasts were able to solubilize low-soluble P sources (Ca and Fe phosphates) and accumulate polyphosphates (polyPs). Sequence analysis of 18S ribosomal deoxyribonucleic acid showed that the yeasts belong to the genera Rhodotorula or Rhodosporidium (EndoGm1) and Cryptococcus (EndoGm11). Results from inoculation experiments showed an effect of the spore-associated yeasts on the root growth of rice, suggesting potential tripartite interactions with mycorrhizal fungi and plants.     

α-Mannosidases of generaAspergillus andRhizopus. Activity and capacity to utilizeSaccharomyces cerevisiae mannan of the best α-mannosidase producerAspergillus flavus link 69

Strains of fungi imperfecti of generaAspergillus andRhizopus were tested for the ability to produce α-mannosidases. The most suitable α-mannosidase producer of a total of 20 strains under study wasAspergillus Ravus Link 69. The parameters studied during the cultivation included the growth rate expressed as cell dry weight, α-mannosidase activity of the extracellular medium withp-nitrophenyl α-D-mannopyranoside as substrate, and utilization ofSaccharomyces cerevisiae mannanvia its disappearance from the cultivation medium.     

Production of extracellular β-mannanases by yeasts and yeast-like microorganisms

A new screening method for simultaneous detection of endo-β-1,4-mannanase and endo-β-1,4-xylanase producing microorganisms is described. Two differently dyed substrate Ostazin Brilliant Red-galactomannan and Remazol Brilliant Blue-xylan were incorporated into the same agar media. Decolorizing of one or both substrates around the cell colonies indicates secretion of the corresponding enzyme(s). The method was used to screen 449 yeasts and yeast-like microorganisms belonging to 68 different genera. The secretion of endo-β-1,4-mannanases and/or endo-β-1,4-xylanases was found within 10 genera (42 positive strains out of 261 tested). A low frequency of occurrence of endo-β-1,4-mannanases was observed within the generaCryptococcus (1 positive strain out of 15 tested),Geotrichum (1 of 6) andPichia (1 of 35). The highest frequency of occurrence of endo-β-1,4-mannanases was found within the generaStephanoascus (2 of 2) andAureobasidium (14 of 14). Strains hydrolyzing Ostazin Brilliant Red-galactomannan were cultivated in liquid media containing 1 % locust bean gum. The best producers of extracellular endo-β-1,4-mannanases were found to be the strains ofAureobasidium pullulans.     

Basidiomycetous Yeasts from Boletales Fruiting Bodies and Their Interactions with the Mycoparasite <Emphasis Type="Italic">Sepedonium chrysospermum</Emphasis> and the Host Fungus <Emphasis Type="Italic">Paxillus</Emphasis>

Interactions between mushrooms, yeasts, and parasitic fungi are probably common in nature, but are rarely described. Bolete fruiting bodies are associated with a broad spectrum of microorganisms including yeasts, and they are commonly infected with filamentous mycoparasites of the genus Sepedonium (teleomorph Hypomyces). We report the isolation of 17 yeast strains from Paxillus and Xerocomus, 16 of which were obtained from the surface tissue, the primary site of Sepedonium infection. Phylogenetic analyses with the D1/D2 region of the 28S ribosomal gene and the internal transcribed spacers placed the yeasts as Rhodotorula, Rhodosporidium, and Mastigobasidium from the Pucciniomycotina, Cryptococcus, Cystofilobasidium, Holtermanniella, and Trichosporon from the Agaricomycotina, and Kluyveromyces from the Saccharomycotina including the first isolation of Rhodotorula graminis from Europe. To investigate the influence of the yeast strains on the mycoparasite and the host fungus, in vitro assays were conducted with Sepedonium chrysospermum and Paxillus involutus. Both S. chrysospermum growth inhibitory and stimulating yeast strains were detected among the isolates. The number of S. chrysospermum inhibitory yeast strains increased and the number of S. chrysospermum stimulatory yeast strains decreased in the presence of P. involutus in co-cultures. Low nutrient levels in the culture medium also led to an increased number of S. chrysospermum inhibitory yeast strains and ten yeasts inhibited the mycoparasite in spatial separation by a crosswall. Six yeast strains inhibited P. involutus in dual culture, and the inhibitory P. involutus yeast interactions increased to nine in the presence of S. chrysospermum. Our results suggest that the bolete-associated yeasts influence the growth of the mycoparasitic fungus, which may affect the health of the fruiting bodies.     

Molecular evolution in yeast of biotechnological interest

The importance of yeast in the food and beverage industries was only realized about 1860, when the role of these organisms in food manufacture became evident. Since they grow on a wide range of substrates and can tolerate extreme physicochemical conditions, yeasts, especially the genera Saccharomyces and Kluyveromyces, have been applied to many industrial processes, Industrial strains of these genera are highly specialized organisms that have evolved to utilize a range of environments and ecological niches to their full potential. This adaptation is called "domestication". This review describes the phylogenetic relationships among Saccharomyces and Kluyveromyces species and the different mechanisms involved in the adaptive evolution of industrial yeast strains.     

Phylogenetic placement of the basidiomycetous yeastsKondoa malvinella andRhodosporidium dacryoidum,and the anamorphic yeastSympodiomycopsis paphiopedili by means of 18S rRNA gene sequence analysis

The 18S ribosomal RNA gene sequences of the basidiomycetous yeastsKondoa malvinella andRhodosporidium dacryoidum, and an anamorphic yeastSympodiomycopsis paphiopedili were determined. The 18S rRNA gene ofR. dacryodium IAM 13522 (ex type) revealed the presence of an intron-like region with a length of 404 nucleotides, which is presumably assigned to a group I intron. The phylogenetic tree, including 34 published reference sequences, was inferred from 1493 sites which could be unambiguously aligned. The molecular phylogeny, using the ascomycetes as an outgroup, divided the basidiomycetes into three major lineages. The first lineage was composed of the smut fungi (Ustilaginales), represented byUstilago maydis, U. hordei, andTilletia caries, includingS. paphiopedili. The second lineage included the type species of teliospore-forming yeast generaLeucosporidium, Rhodosporidium, andSporidiobolus, and the generaErythrobasidium andKondoa, both previously included in the Filobasidiaceae.Rhodosporidium dacryodium showed a close relationship withE. hasegawianum, which was backed by a high bootstrap support. The rust fungiCronartium ribicola andPeridermium harknessii were also included in this lineage. The last lineage was formed by the filobasidiaceous yeasts,Cystofilobasidium capitatum, Mrakia frigida, Filobasidium floriforme, andFilobasidiella neoformans, and the anamorphic yeastsBullera alba (the anamorph ofBulleromyces albus) andTrichosporon cutaneum. Members ofTremella and selected hymenomycetous genera were also included in this lineage.The nucleotide sequence data reported in this paper will appear in the GSDB, DDBJ, EMBL and NCBI nucleotide sequence databases with the following accession numbers D13459 (Rhodosporidium dacryodium), D13776 (Kondoa malvinella), and D14006 (Sympodiomycopsis paphiopedili).     

New search for pectolytic yeasts

A new screening method for pectin-depolymerizing microorganisms is described. The method is based on precipitation of non-hydrolyzed citrus pectin with hexadecyltrimethylammonium bromide in a medium solidified with a bacterial gelling gum. A substrate depolymerized by the secreted enzymes does not precipitate, and the positive strains thus show transparent areas around the colonies. The method was used to screen 300 yeast and yeast-like microorganisms belonging to 52 different genera. The secretion of pectin-depolymerizing enzymes occured with different frequencies in 13 genera (69 positive strains of 207 tested), the lowest frequency being found in the genusCandida (13 positive out of 125 strains tested) and the highest frequency in the generaAureobasidium (4 of 6)Cryptococcus (29 of 38),Geotrichum (4 of 9),Kluyveromyces (5 of 5),Rhodosporidium (2 of 2),Leucosporidium (2 of 2),Trichosporon (3 of 6) andUstilago (2 of 2). Strains giving the highest number of harvested cells after growth on pectin in a liquid medium have been identified. Supported by theGrant Agency for Science of the Slovak Republic.     

Cloning, characterization and heterologous expression of epoxide hydrolase-encoding cDNA sequences from yeasts belonging to the genera Rhodotorula and Rhodosporidium

Epoxide hydrolase-encoding cDNA sequences were isolated from the basidiomycetous yeast species Rhodosporidium toruloides CBS 349, Rhodosporidium toruloides CBS 14 and Rhodotorula araucariae CBS 6031 in order to evaluate the molecular data and potential application of this type of enzymes. The deduced amino acid sequences were similar to those of the known epoxide hydrolases from Rhodotorula glutinis CBS 8761, Xanthophyllomyces dendrorhous CBS 6938 and Aspergillus niger LCP 521, which all correspond to the group of the microsomal epoxide hydrolases. The epoxide hydrolase encoding cDNAs of the Rhodosporidium and Rhodotorula species were expressed in Escherichia coli. The recombinant strains were able to hydrolyze trans-1-phenyl-1,2-epoxypropane with high enantioselectivity.     

The adventures of the yeast genusEndomycopsis Dekker

The yeast generaEndomyces, Endomycopsella, Guilliermondella andSaccharomycopsis are delimited by the size, structure and pigmentation of the ascospores; they include mycelial yeasts formerly classified in the invalid genusEndomycopsis. The ultrastructure of the cell wall and the septa of yeasts is briefly discussed.