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1.
Extremophiles ; 28(1): 17, 2024 Feb 11.
Article in English | MEDLINE | ID: mdl-38342818

ABSTRACT

Yeasts from cold environments have a wide range of strategies to prevent the negative effects of extreme conditions, including the production of metabolites of biotechnological interest. We investigated the growth profile and production of metabolites in yeast species isolated from cold environments. Thirty-eight strains were tested for their ability to grow at different temperatures (5-30 °C) and solute concentrations (3-12.5% NaCl and 50% glucose). All strains tested were able to grow at 5 °C, and 77% were able to grow with 5% NaCl at 18 °C. We were able to group strains based on different physicochemical/lifestyle profiles such as polyextremotolerant, osmotolerant, psychrotolerant, or psychrophilic. Five strains were selected to study biomass and metabolite production (glycerol, trehalose, ergosterol, and mycosporines). These analyses revealed that the accumulation pattern of trehalose and ergosterol was related to each lifestyle profile. Also, our findings would suggest that mycosporines does not have a role as an osmolyte. Non-conventional fermentative yeasts such as Phaffia tasmanica and Saccharomyces eubayanus may be of interest for trehalose production. This work contributes to the knowledge of non-conventional yeasts with biotechnological application from cold environments, including their growth profile, metabolites, and biomass production under different conditions.


Subject(s)
Basidiomycota , Trehalose , Trehalose/metabolism , Sodium Chloride/metabolism , Yeasts , Ergosterol/metabolism , Cold Temperature
2.
Int J Mol Sci ; 24(6)2023 Mar 21.
Article in English | MEDLINE | ID: mdl-36983003

ABSTRACT

The Phaffia rhodozyma UCD 67-385 genome harbors a 7873 bp cluster containing DDGS, OMT, and ATPG, encoding 2-desmethy-4-deoxygadusol synthase, O-methyl transferase, and ATP-grasp ligase, respectively, of the mycosporine glutaminol (MG) biosynthesis pathway. Homozygous deletion mutants of the entire cluster, single-gene mutants, and the Δddgs-/-;Δomt-/- and Δomt-/-;Δatpg-/- double-gene mutants did not produce mycosporines. However, Δatpg-/- accumulated the intermediate 4-deoxygadusol. Heterologous expression of the DDGS and OMT or DDGS, OMT, and ATPG cDNAs in Saccharomyces cerevisiae led to 4-deoxygadusol or MG production, respectively. Genetic integration of the complete cluster into the genome of the non-mycosporine-producing CBS 6938 wild-type strain resulted in a transgenic strain (CBS 6938_MYC) that produced MG and mycosporine glutaminol glucoside. These results indicate the function of DDGS, OMT, and ATPG in the mycosporine biosynthesis pathway. The transcription factor gene mutants Δmig1-/-, Δcyc8-/-, and Δopi1-/- showed upregulation, Δrox1-/- and Δskn7-/- showed downregulation, and Δtup6-/- and Δyap6-/- showed no effect on mycosporinogenesis in glucose-containing medium. Finally, comparative analysis of the cluster sequences in several P. rhodozyma strains and the four newly described species of the genus showed the phylogenetic relationship of the P. rhodozyma strains and their differentiation from the other species of the genus Phaffia.


Subject(s)
Basidiomycota , Phylogeny , Homozygote , Sequence Deletion , Basidiomycota/genetics , Saccharomyces cerevisiae
3.
FEMS Yeast Res ; 21(1)2021 01 22.
Article in English | MEDLINE | ID: mdl-33232451

ABSTRACT

Cold environments impose challenges to organisms. Polyextremophile microorganisms can survive in these conditions thanks to an array of counteracting mechanisms. Naganishia vishniacii, a yeast species hitherto only isolated from McMurdo Dry Valleys, Antarctica, is an example of a polyextremophile. Here we present the first draft genomic sequence of N. vishniacii. Using comparative genomics, we unraveled unique characteristics of cold associated adaptations. 336 putative genes (total: 6183) encoding solute transfers and chaperones, among others, were absent in sister species. Among genes shared by N. vishniacii and its closest related species we found orthologs encompassing possible evidence of positive selection (dN/dS > 1). Genes associated with photoprotection were found in agreement with high solar irradiation exposure. Also genes coding for desaturases and genomic features associated with cold tolerance (i.e. trehalose synthesis and lipid metabolism) were explored. Finally, biases in amino acid usage (namely an enrichment of glutamine and a trend in proline reduction) were observed, possibly conferring increased protein flexibility. To the best of our knowledge, such a combination of mechanisms for cold tolerance has not been previously reported in fungi, making N. vishniacii a unique model for the study of the genetic basis and evolution of cold adaptation strategies.


Subject(s)
Adaptation, Physiological/genetics , Basidiomycota/genetics , Cold Temperature , Genome, Microbial , Antarctic Regions , Evolution, Molecular , Genomics/methods
4.
Methods Mol Biol ; 1852: 297-310, 2018.
Article in English | MEDLINE | ID: mdl-30109639

ABSTRACT

Astaxanthin is a xanthophyll pigment of high economic value for its use as a feeding component in aquaculture. Phaffia rhodozyma (Xanthophyllomyces dendrorhous) is a basidiomycetous fungi able to synthesize astaxanthin as its major carotenoid, the only known yeast species bearing the capability to produce this type of carotenoid and the only tremellomycetes with biotechnological application. Recently, the habitat and intraspecific variability of this species have been found to be wider than previously expected, encouraging the search for new wild strains with potential biotechnological applications. Here we describe effective procedures for isolation of P. rhodozyma from environmental samples, accurate identification of the strains, analysis of their astaxanthin content, and proper conservation of the isolates.


Subject(s)
Basidiomycota/isolation & purification , Basidiomycota/metabolism , Basidiomycota/genetics , Conjugation, Genetic , DNA, Ribosomal Spacer , Environment , Fermentation , Glucose/metabolism , Phenotype , Spectrum Analysis , Starch/chemistry , Starch/metabolism , Xanthophylls/biosynthesis , Xanthophylls/chemistry , Xanthophylls/isolation & purification
5.
BMC Genomics ; 17(1): 901, 2016 11 09.
Article in English | MEDLINE | ID: mdl-27829365

ABSTRACT

BACKGROUND: The class Tremellomycete (Agaricomycotina) encompasses more than 380 fungi. Although there are a few edible Tremella spp., the only species with current biotechnological use is the astaxanthin-producing yeast Phaffia rhodozyma (Cystofilobasidiales). Besides astaxanthin, a carotenoid pigment with potent antioxidant activity and great value for aquaculture and pharmaceutical industries, P. rhodozyma possesses multiple exceptional traits of fundamental and applied interest. The aim of this study was to obtain, and analyze two new genome sequences of representative strains from the northern (CBS 7918T, the type strain) and southern hemispheres (CRUB 1149) and compre them to a previously published genome sequence (strain CBS 6938). Photoprotection and antioxidant related genes, as well as genes involved in sexual reproduction were analyzed. RESULTS: Both genomes had ca. 19 Mb and 6000 protein coding genes, similar to CBS 6938. Compared to other fungal genomes P. rhodozyma strains and other Cystofilobasidiales have the highest number of intron-containing genes and highest number of introns per gene. The Patagonian strain showed 4.4 % of nucleotide sequence divergence compared to the European strains which differed from each other by only 0.073 %. All known genes related to the synthesis of astaxanthin were annotated. A hitherto unknown gene cluster potentially responsible for photoprotection (mycosporines) was found in the newly sequenced P. rhodozyma strains but was absent in the non-mycosporinogenic strain CBS 6938. A broad battery of enzymes that act as scavengers of free radical oxygen species were detected, including catalases and superoxide dismutases (SODs). Additionally, genes involved in sexual reproduction were found and annotated. CONCLUSIONS: A draft genome sequence of the type strain of P. rhodozyma is now available, and comparison with that of the Patagonian population suggests the latter deserves to be assigned to a distinct variety. An unexpected genetic trait regarding high occurrence of introns in P. rhodozyma and other Cystofilobasidiales was revealed. New genomic insights into fungal homothallism were also provided. The genetic basis of several additional photoprotective and antioxidant strategies were described, indicating that P. rhodozyma is one of the fungi most well-equipped to cope with environmental oxidative stress, a factor that has probably contributed to shaping its genome.


Subject(s)
Basidiomycota/genetics , Genetic Variation , Genome, Fungal , Genomics , Antioxidants/metabolism , Basidiomycota/metabolism , Basidiomycota/radiation effects , Catalase/metabolism , Computational Biology/methods , Gene Order , Genomics/methods , High-Throughput Nucleotide Sequencing , Reproduction, Asexual/genetics , Ultraviolet Rays
6.
Recent Pat Biotechnol ; 8(3): 179-93, 2014.
Article in English | MEDLINE | ID: mdl-25619303

ABSTRACT

Exposure to ultraviolet radiation (UVR) is harmful to living organisms, causing damage to macromolecules such as DNA, RNA, proteins and lipids. Depending on the wavelength, the injury could be direct or indirect through reactive oxygen intermediates, so it is desirable to find compounds that can reduce both. Many organic chemicals used in commercial sunscreen possess estrogenic activity in vivo. In this report we analyzed recent patents related to UV sunscreens of microbial origin, in particular mycosporines (MYC) and mycosporine-like aminoacids (MAA). Both are promising natural alternatives for both direct (UV-absorption) and indirect (antioxidant) protection, given they show strong photostability and absence of cytotoxicity. It becomes clear that although the search for natural photoprotective molecules is relatively recent, efforts have been invested mainly in marine environments, remaining still many potential photoprotective molecules to find in other type of habitats. Furthermore, unicellular microorganisms have several advantages for the production of metabolites of interest, since they improve the production costs due to its simplicity of culture and easy genetic manipulation. The knowledge of the biosynthesis pathway of MYC and MAA is essential to improve rationally their expression levels. Currently, only the MAA pathway in bacteria has been reported, remaining the MYC pathway unclear. Future perspectives include the heterologous expression of MYC and/or MAA in industrially friendly microorganisms (bacteria and yeast) in order to co-produce different UV-protective molecules and thus cover a broader UV spectrum and simplify the production process.


Subject(s)
Amino Acids/metabolism , Bacteria/metabolism , Protective Agents/metabolism , Sunscreening Agents/metabolism , Yeasts/metabolism , Animals , Biosynthetic Pathways/physiology , Humans , Patents as Topic , Ultraviolet Rays
7.
J Basic Microbiol ; 53(9): 766-72, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23417292

ABSTRACT

Xanthophyllomyces dendrorhous (Phaffia rhodozyma) yeasts are biotechnologically exploited as a natural source of astaxanthin for aquaculture. Based on results of recent studies, it has become clear that this species possesses a greater genetic variability generating the necessity to uncover it and assess its potential for the astaxanthin industry. However, difficulties for the isolation of the X. dendrorhous hinder extensive environmental surveys which need to be carried out to better understand the habitat, distribution and genetic diversity of this species. We extensively searched for distinctive physiological traits of X. dendrorhours by testing phenotypic properties simultaneously with a panel of common sympatric fungi. As a result we obtained a new and innovative strategy for improving X. dendrorhous recovery rate and identification from environmental samples. This strategy involved the use of trehalose-based media, and a rapid X. dendrorhous identification method based on the simultaneous spectrophotometric detection of astaxanthin and UV-absorbing compounds (mycosporines). The proposed procedures proved effective in field trials conducted in natural environments of Patagonia (Argentina) and thus represent an important tool for the discovery of new astaxanthin-producing strains of X. dendrorhous useful for the aquaculture industry.


Subject(s)
Basidiomycota/isolation & purification , Basidiomycota/metabolism , Environmental Microbiology , Mycology/methods , Aquaculture/methods , Argentina , Culture Media/chemistry , Spectrophotometry , Xanthophylls/metabolism
8.
Methods Mol Biol ; 898: 183-94, 2012.
Article in English | MEDLINE | ID: mdl-22711126

ABSTRACT

Astaxanthin is a xanthophyll pigment of high economic value for its use as a feeding component in aquaculture. Xanthophyllomyces dendrorhous is a basidiomycetous fungi able to synthesize astaxanthin as its major carotenoid, and the only known yeast species bearing the capability to produce this type of carotenoid. Recently, the habitat and intraspecific variability of this species have been found to be wider than previously expected, encouraging the search for new wild strains with potential biotechnological applications. Here we describe effective procedures for isolation of X. dendrorhous from environmental samples, accurate identification of the strains, analysis of their astaxanthin content, and proper conservation of the isolates.


Subject(s)
Basidiomycota/isolation & purification , Basidiomycota/metabolism , Environment , Fermentation , Glucose/metabolism , Histocytological Preparation Techniques , Phenotype , Plant Leaves/microbiology , Starch/biosynthesis , Water Microbiology , Xanthophylls/analysis , Xanthophylls/biosynthesis , Xanthophylls/isolation & purification
9.
Methods Mol Biol ; 898: 275-83, 2012.
Article in English | MEDLINE | ID: mdl-22711133

ABSTRACT

Yeasts of the genera Rhodotorula are able to synthesize different pigments of high economic value like ß-carotene, torulene, and torularhodin, and therefore represent a biotechnologically interesting group of yeasts. However, the low production rate of pigment in these microorganisms limits its industrial application. Here we describe some strategies to obtain hyperpigmented mutants of Rhodotorula mucilaginosa by means of ultraviolet-B radiation, the procedures for total carotenoids extraction and quantification, and a method for identification of each pigment.


Subject(s)
Carotenoids/biosynthesis , Metabolic Engineering/methods , Rhodotorula/genetics , Rhodotorula/metabolism , Carotenoids/analysis , Carotenoids/isolation & purification , Chromatography, High Pressure Liquid , Mutagenesis , Mutation , beta Carotene/analysis , beta Carotene/biosynthesis , beta Carotene/isolation & purification
10.
Braz. j. microbiol ; 42(3): 937-947, July-Sept. 2011. tab
Article in English | LILACS | ID: lil-607522

ABSTRACT

The diversity of yeasts collected from different sites in Antarctica (Admiralty Bay, King George Island and Port Foster Bay and Deception Island) and their ability to produce extracellular enzymes and mycosporines were studied. Samples were collected during the austral summer season, between November 2006 and January 2007, from the rhizosphere of Deschampsia antarctica, ornithogenic (penguin guano) soil, soil, marine and lake sediments, marine water and freshwater from lakes. A total of 89 isolates belonging to the following genera were recovered: Bensingtonia, Candida, Cryptococcus, Debaryomyces, Dioszegia, Exophiala, Filobasidium, Issatchenkia (Pichia), Kodamaea, Leucosporidium, Leucosporidiella, Metschnikowia, Nadsonia, Pichia, Rhodotorula, and Sporidiobolus, and the yeast-like fungi Aureobasidium, Leuconeurospora and Microglossum. Cryptococcus victoriae was the most frequently identified species. Several species isolated in our study have been previously reported to be Antarctic psychophilic yeasts, including Cr. antarcticus, Cr. victoriae, Dioszegia hungarica and Leucosporidium scottii. The cosmopolitan yeast species A. pullulans, C. zeylanoides, D. hansenii, I. orientalis, K. ohmeri, P. guilliermondii, Rh. mucilaginosa, and S. salmonicolor were also isolated. Five possible new species were identified. Sixty percent of the yeasts had at least one detectable extracellular enzymatic activity. Cryptococcus antarcticus, D. aurantiaca, D. crocea, D. hungarica, Dioszegia sp., E. xenobiotica, Rh. glaciales, Rh. laryngis, Microglossum sp. 1 and Microglossum sp. 2 produced mycosporines. Of the yeast isolates, 41.7 percent produced pigments and/or mycosporines and could be considered adapted to survive in Antarctica. Most of the yeasts had extracellular enzymatic activities at 4ºC and 20ºC, indicating that they could be metabolically active in the sampled substrates.


Subject(s)
Biodiversity , Environmental Microbiology , Enzyme Activation , Enzymes/analysis , Yeasts/isolation & purification , Yeasts/metabolism , Rhizophoraceae/genetics , Rhizophoraceae/metabolism , Seawater , Methods , Methods
11.
Yeast ; 28(8): 619-27, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21744380

ABSTRACT

The synthesis of the UV Absorbing compounds named mycosporines (MYCs) has been detected in a few basidiomycetous yeast groups. Conspicuous accumulation of mycosporine-glutaminol-glucoside (MGG) in yeasts requires photo-induction and its photoprotective function has been postulated. The distribution of the ability to produce MYCs appeared to be related to the yeast taxonomic affiliation. In view of the potential significance of MYCs in yeast taxonomy, we here studied the distribution of this trait among dimorphic basidiomycetes of the Pucciniomycotina. Of the 94 fungal species (377 strains and 33 genera) tested, almost half were MYC-positive and MGG was the main compound produced. MGG synthesis was observed for representatives of five of seven Pucciniomycotina classes, indicating that this trait is widely distributed in this group. MGG detection proved useful for the differentiation of species of the polyphyletic genera, such as Rhodotorula and Sporobolomyces, that are phylogenetically separated. MGG quantification and UV tolerance studies in Cystobasidiomycetes supported the idea that the habitat of origin of each strains is important in the level of MGG synthesis and that MYCs have a photoprotective function in yeasts. The taxonomic value of this trait in fungal systematics is discussed.


Subject(s)
Basidiomycota/classification , Basidiomycota/metabolism , Cyclohexanols/metabolism , Glucosides/metabolism , Phylogeny , Basidiomycota/genetics , Basidiomycota/radiation effects , Molecular Sequence Data , Ultraviolet Rays
12.
FEMS Microbiol Ecol ; 76(1): 1-13, 2011 Apr.
Article in English | MEDLINE | ID: mdl-21223324

ABSTRACT

Nahuel Huapi (NH) Lake is an oligotrophic temperate lake of glacial origin with high transparency, surrounded by well-developed forests and located at San Carlos de Bariloche, Nahuel Huapi National Park, in Patagonia, Argentina. In this lake, we characterized yeast distribution and diversity along a south-to-north transect and established a relationship between the ability to produce photoprotective compounds (PPCs) (carotenoid pigments and mycosporines) and the occurrence of yeast at different collection points. Subsurface water samples were filtered for yeast isolation. Total yeast counts ranged between 22 and 141 CFU L(-1) , and the highest values corresponded to the most impacted sites. Littoral sites had a low proportion of yeast-producing PPCs and this group prevailed in pelagic sites. This is probably a result of the high transparency of the water and the increased UV exposure. The yeast community from NH Lake showed a high species richness and a uniform distribution of taxa between pelagic and border collection points. Yeasts were identified as belonging to 14 genera and 34 species. Rhodotorula mucilaginosa and Cryptococcus victoriae were the most frequently found species, representing 14.4% and 13.6% of the total yeast isolates, respectively. Most of the yeast isolates demonstrated at least one extracellular enzymatic activity (mainly cellulase and lipase activities), which suggested that these microorganisms are metabolically active in the lake.


Subject(s)
Biodiversity , Cryptococcus/isolation & purification , Rhodotorula/isolation & purification , Water Microbiology , Amino Acids/analysis , Argentina , Carotenoids/analysis , Cellulase/analysis , Colony Count, Microbial , Cryptococcus/classification , Cryptococcus/enzymology , DNA Fingerprinting , DNA, Fungal/genetics , Lipase/analysis , Rhodotorula/classification , Rhodotorula/enzymology , Sequence Analysis, DNA
13.
FEMS Yeast Res ; 11(1): 52-9, 2011 Feb.
Article in English | MEDLINE | ID: mdl-20955516

ABSTRACT

The ability of the basidiomycetous yeast Xanthophyllomyces dendrorhous (Phaffia rhodozyma) to accumulate astaxanthin is responsible for the industrial use of this yeast as a microbial source of pigments for aquaculture. It is also hypothesized that astaxanthin accounts for its ability to thrive in highly oxidative and UV-exposed habitats. Here, we assessed the ability of this species to synthesize UV-absorbing compounds generally known as mycosporines, evaluated the effect of culture media in the production of these compounds and compared its UV growth resistance and tolerance with other yeasts. The 48 wild and collection strains screened were positive for mycosporines and a unique compound identified as mycosporine-glutaminol-glucoside (MGG) was detected. Thus, the ability of X. dendrorhous to produce MGG, as described here for the first time, is so far unique among the Cystofilobasidiales. The compound was synthesized constitutively, although growth under visible light and, to a greater extent, UVA radiation stimulated its production. Strains from UV-exposed habitats produced larger quantities and oligotrophic complex media seemed to favor MGG accumulation. UV tolerance and survival of X. dendrorhous was high and comparable to that of the polyextremophilic Rhodotorula mucilaginosa. The taxonomical and ecological implications of the production of MGG by X. dendrorhous are discussed.


Subject(s)
Basidiomycota/metabolism , Basidiomycota/radiation effects , Cyclohexanols/metabolism , Glucosides/metabolism , Basidiomycota/growth & development , Culture Media/chemistry , Microbial Viability/radiation effects , Ultraviolet Rays
14.
Radiat Res ; 175(1): 44-50, 2011 Jan.
Article in English | MEDLINE | ID: mdl-21175346

ABSTRACT

Several yeast species are able to synthesize and accumulate UV-radiation-absorbing mycosporine metabolites that are of unclear physiological function. In this work we analyzed the relationship between mycosporine-glutaminol glucoside (MGG) production, cell survival after UVB irradiation, and formation of cyclobutane pyrimidine dimers (CPDs). We also assessed the photostability and singlet oxygen quenching activity of MGG. A set of nine isolates of the basidiomycetous yeast Cryptococcus steppossus cultured in both dark and light conditions was used for the studies. Survival of the UVB-irradiated isolates and MGG concentration had a linear relationship when the concentration was over 2.5 mg g(-1). CPD accumulation and MGG accumulation were inversely related. MGG in aqueous solution was photostable with a photodecomposition quantum yield of 1.16 × 10(-5). MGG quenching of singlet oxygen was also observed, and the rate constant for the process in D(2)O was 5.9 × 10(7) M(-1) s(-1). Our results support the idea that MGG plays an important role as a UVB photoprotective metabolite in yeasts by protecting against direct damage on DNA and probably against indirect damage by singlet oxygen quenching.


Subject(s)
Antioxidants/metabolism , Cryptococcus/radiation effects , Cyclohexanols/metabolism , Glucosides/metabolism , Ultraviolet Rays , Cryptococcus/metabolism , Cyclohexanols/chemistry , DNA Damage , Glucosides/chemistry , Pyrimidine Dimers/metabolism , Singlet Oxygen/metabolism
15.
Braz J Microbiol ; 42(3): 937-47, 2011 Jul.
Article in English | MEDLINE | ID: mdl-24031709

ABSTRACT

The diversity of yeasts collected from different sites in Antarctica (Admiralty Bay, King George Island and Port Foster Bay and Deception Island) and their ability to produce extracellular enzymes and mycosporines were studied. Samples were collected during the austral summer season, between November 2006 and January 2007, from the rhizosphere of Deschampsia antarctica, ornithogenic (penguin guano) soil, soil, marine and lake sediments, marine water and freshwater from lakes. A total of 89 isolates belonging to the following genera were recovered: Bensingtonia, Candida, Cryptococcus, Debaryomyces, Dioszegia, Exophiala, Filobasidium, Issatchenkia (Pichia), Kodamaea, Leucosporidium, Leucosporidiella, Metschnikowia, Nadsonia, Pichia, Rhodotorula, and Sporidiobolus, and the yeast-like fungi Aureobasidium, Leuconeurospora and Microglossum. Cryptococcus victoriae was the most frequently identified species. Several species isolated in our study have been previously reported to be Antarctic psychophilic yeasts, including Cr. antarcticus, Cr. victoriae, Dioszegia hungarica and Leucosporidium scottii. The cosmopolitan yeast species A. pullulans, C. zeylanoides, D. hansenii, I. orientalis, K. ohmeri, P. guilliermondii, Rh. mucilaginosa, and S. salmonicolor were also isolated. Five possible new species were identified. Sixty percent of the yeasts had at least one detectable extracellular enzymatic activity. Cryptococcus antarcticus, D. aurantiaca, D. crocea, D. hungarica, Dioszegia sp., E. xenobiotica, Rh. glaciales, Rh. laryngis, Microglossum sp. 1 and Microglossum sp. 2 produced mycosporines. Of the yeast isolates, 41.7% produced pigments and/or mycosporines and could be considered adapted to survive in Antarctica. Most of the yeasts had extracellular enzymatic activities at 4°C and 20°C, indicating that they could be metabolically active in the sampled substrates.

16.
Photochem Photobiol Sci ; 9(8): 1145-51, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20571712

ABSTRACT

In this paper we report the relationship between carotenoids and ergosterol and cell UV-B resistance in different strains of the yeast Rhodotorula mucilaginosa. Cell survival was studied using a set of 13 strains; additionally, two mutants (a hyper-producing one and a colourless one) in combination with diphenylamine (DPA), a carotenogenesis inhibitor, were used. A positive correlation between total carotenoids and survival to UV-B radiation was found. However, when individual carotenoid concentrations were tested, only torularhodin was found to be significantly related to UV-B survival. On the contrary, ergosterol did not affect survival. The hyper-pigmented strain showed an enhanced survival (up to 250%) compared to the parental strain, while the survival of the albino mutant was similar to that experienced by the parental strain; however, observed changes in survival were dose dependent. The cyclobutane pyrimidine dimers (CPDs), one of the major forms of DNA damage caused by UV exposure, appears as unrelated to the accumulation of carotenoids and cell survival. These results indicate that bearing higher torularhodin concentrations enhances UV-B survival in yeasts and, thus, the accumulation of this pigment constitutes an important mechanism that improves the resistance of yeasts to UV-B.


Subject(s)
Carotenoids/physiology , Rhodotorula/radiation effects , Carotenoids/pharmacology , Cell Survival , DNA Damage , Diphenylamine/pharmacology , Ergosterol/pharmacology , Pyrimidine Dimers/chemistry , Rhodotorula/drug effects , Ultraviolet Rays
17.
FEMS Microbiol Ecol ; 69(3): 353-62, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19624739

ABSTRACT

Mountain lakes located at a high elevation are typically exposed to high UV radiation (UVR). Little is known about the ecology and diversity of yeasts inhabiting these extreme environments. We studied yeast occurrence (with special emphasis on those producing carotenoid pigments) at five high-altitude (>1400 m a.s.l.) water bodies located in the Nahuel Huapi National Park (Bariloche, Argentina). Isolates were identified using a polyphasic approach. Production of photoprotective compounds (carotenoids and mycosporines) by yeast isolates, and UVB resistance of selected species were studied. All water samples contained viable yeast cells in variable numbers, generally ranging from 49 to 209 cells L(-1). A total of 24 yeast species was found; at least four represented novel species. Carotenogenic yeasts prevailed in lakes with low water conductivity and higher transparency and chlorophyll a levels. Apparently, the ability to produce photoprotective compounds in yeasts was related to the transparency of mountain lake waters, and strains from more transparent waters developed increased UVB resistance. Our results indicate that UVR is an important environmental factor affecting the yeast community structure in aquatic habitats.


Subject(s)
Fresh Water/microbiology , Ultraviolet Rays , Water Microbiology , Yeasts/radiation effects , Argentina , Biodiversity , DNA, Fungal/genetics , RNA, Ribosomal/genetics , Yeasts/classification , Yeasts/genetics , Yeasts/isolation & purification
18.
J Photochem Photobiol B ; 95(3): 156-61, 2009 Jun 03.
Article in English | MEDLINE | ID: mdl-19342251

ABSTRACT

In this work, the photoprotective role of carotenoids in yeasts was analysed by contrasting the responses to UV-B of pigmented and naturally occurring albino strains of Sporobolomyces ruberrimus and Cystofilobasidium capitatum in different conditions. Albino and pigmented strains were confirmed to be conspecific by PCR fingerprinting and rDNA sequencing. Experimental exposure to UV-B conducted with both yeast species showed that the pigmented strains were more tolerant to UV-B than the albino strains and that the increment in carotenoid contents during the stationary growth phase enhance survivorship. These results indicated that carotenoid pigments afford UV-B protection in yeasts.


Subject(s)
Basidiomycota/radiation effects , Carotenoids/biosynthesis , Ultraviolet Rays , Basidiomycota/growth & development , Basidiomycota/metabolism , Cell Survival , Pigmentation
19.
J Ind Microbiol Biotechnol ; 35(3): 151-8, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18000694

ABSTRACT

A novel population of the biotechnologically important yeast Xanthophyllomyces dendrorhous, the sexual stage of Phaffia rhodozyma, has been recently isolated for the first time in the southern Hemisphere (Patagonia, Argentina). The aim of the present work was to phenotypically and genotypically characterize two representative strains of this new population, and assess such strains as a potential biotechnological source of astaxanthin, fatty acids and extracellular enzymes. Minor variations were found in physiological tests. PCR fingerprinting studies (MSP-PCR) showed the main differences between X. dendrorhous Patagonian and Type strains. Patagonian strains accumulated a xanthophyll-like pigment, which was identified as astaxanthin. These strains showed low fatty acids content (mainly polyunsaturated fatty acids) and, of a total of six extracellular enzymes tested, only produced amylase. Genetic differences between Patagonian and collection X. dendrorhous strains could be explained by geographic isolation and habitat specificity.


Subject(s)
Basidiomycota/classification , Basidiomycota/metabolism , Phenotype , Amylases/isolation & purification , Argentina , Basidiomycota/genetics , Culture Media , DNA Fingerprinting , DNA, Fungal , Fatty Acids, Unsaturated/isolation & purification , Genotype , Mycological Typing Techniques , Species Specificity , Xanthophylls/biosynthesis
20.
Photochem Photobiol ; 82(4): 972-80, 2006.
Article in English | MEDLINE | ID: mdl-17205625

ABSTRACT

In this paper we present the results of research on the occurrence, induction and role of photoprotective compounds (PPCs) present in native aquatic yeasts from freshwater Patagonian ecosystems. We focus on the effect of UV radiation (UVR) as a factor that controls the level of photoprotection of yeasts, and explore its potential significance in shaping yeast distributional patterns. The research presented here combines field surveys and laboratory work, including the isolation and culture of native yeasts strains, and laboratory assays under different radiation conditions. The results obtained suggest that yeasts are common dwellers of oligotrophic Patagonian water bodies, and provide the first evidence of the distribution of PPC (carotenoid and mycosporine)-producing yeasts in temperate freshwaters. A greater proportion of carotenogenic yeasts were observed in high-elevation lakes. The yeast strains isolated from these environments were found to produce higher amounts of mycosporines (MYCs), and to present higher tolerance to UVB exposure than those from piedmont lakes. Patagonian yeasts have only one type of MYC, mycosporine-glutaminol-glucoside (myc-glu-glu), which seems common to all other yeasts. By analyzing the production of myc-glu-glu in a large number of yeasts belonging to different taxonomic groups, we propose that this compound may have potential use as a chemotaxonomic marker in yeast systematics. Collectively, our work reveals that in Patagonian freshwater yeasts there is an apparent relationship between the ability to produce PPCs, their tolerance to UV exposure and their success in colonizing habitats highly exposed to UVR.


Subject(s)
Ecosystem , Fresh Water , Yeasts/radiation effects , Argentina , Carotenoids/biosynthesis , Carotenoids/chemistry , DNA, Fungal/genetics , Molecular Structure , Photochemistry , Phylogeny , Pigments, Biological/metabolism , Ribosomes/genetics , Yeasts/genetics , Yeasts/metabolism
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