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PubMed2003

Hydrocarbon degradation and enzyme activities of cold-adapted bacteria and yeasts.

The potential of 89 culturable cold-adapted isolates from uncontaminated habitats, including 61 bacterial and 28 yeast strains, to utilize representative fractions of petroleum hydrocarbons (n-alkanes, monoaromatic and polycyclic aromatic hydrocarbons) for growth and to produce various enzymes at 10 degrees C was investigated. The efficiency of bacterial and yeast strains was compared. The growth temperature range of the yeast strains was significantly smaller than that of the bacterial strains. Sixty percent of the yeasts but only 8% of the bacteria could be classified as true psychrophiles, showing no growth above 20 degrees C. A high percentage (89%) of the yeast strains showed lipase activity. More than one-third of the 61 bacterial strains produced amylase, beta-lactamase, beta-galactosidase or lipase; more than two-thirds were protease producers. Only 6% of the bacterial strains but 79% of the yeast strains utilized n-hexadecane for growth; 13% of the bacterial strains and 21-32% of the yeast strains utilized phenol, phenanthrene or anthracene for growth. Only four yeast strains but none of the bacterial strains could grow with all hydrocarbons tested. The biodegradation of phenol was investigated in fed-batch cultures at 10 degrees C. Three yeast strains degraded phenol concentrations as high as 10 mM (one strain) or 12.5 mM (two strains). Of eight bacterial strains, two strains degraded up to 10 mM phenol. The optimum temperature for phenol degradation was 20 degrees C for all eight bacterial strains and for two yeast strains. Biodegradation by five yeast strains was optimal at 10 degrees C and faster at 1 degrees C than at 20 degrees C. All phenol-degrading strains produced catechol 1,2 dioxygenase activity.

Adaptation, Physiological↗
PubMed2003

Yeast tRNA(Phe) expressed in human cells can be selected by HIV-1 for use as a reverse transcription primer.

All naturally occurring human immune deficiency viruses (HIV-1) select and use tRNA(Lys,3) as the primer for reverse transcription. Studies to elucidate the mechanism of tRNA selection from the intracellular milieu have been hampered due to the difficulties in manipulating the endogenous levels of tRNA(Lys,3). We have previously described a mutant HIV-1 with a primer binding site (PBS) complementary to yeast tRNA(Phe) (psHIV-Phe) that relies on transfection of yeast tRNA(Phe) for infectivity. To more accurately recapitulate the selection process, a cDNA was designed for the intracellular expression of the yeast tRNA(Phe). Increasing amounts of the plasmid encoding tRNA(Phe) resulted in a corresponding increase in levels of yeast tRNA(Phe) in the cell. The yeast tRNA(Phe) isolated from cells transfected with the cDNA for yeast tRNA(Phe), or in the cell lines expressing yeast tRNA(Phe), were aminoacylated, indicating that the expressed yeast tRNA(Phe) was incorporated into tRNA biogenesis pathways and translation. Increasing the cytoplasmic levels of tRNA(Phe) resulted in increased encapsidation of tRNA(Phe) in viruses with a PBS complementary to tRNA(Phe) (psHIV-Phe) or tRNA(Lys,3) (wild-type HIV-1). Production of infectious psHIV-Phe was dependent on the amount of cotransfected tRNA(Phe) cDNA. Increasing amounts of plasmids encoding yeast tRNA(Phe) produced an increase of infectious psHIV-Phe that plateaued at a level lower than that from the transfection of the wild-type genome, which uses tRNA(Lys,3) as the primer for reverse transcription. Cell lines were generated that expressed yeast tRNA(Phe) at levels approximately 0.1% of that for tRNA(Lys,3). Even with this reduced level of yeast tRNA(Phe), the cell lines complemented psHIV-Phe over background levels. The results of these studies demonstrate that intracellular levels of primer tRNA can have a direct effect on HIV-1 infectivity and further support the role for PBS-tRNA complementarity in the primer selection process.

Amino Acyl-tRNA Synthetases↗
PubMed2002

Yeast genomic expression studies using DNA microarrays.

The exploration and characterization of yeast genomic expression programs is providing a wealth of information about yeast biology, as well as other organisms. The intriguing biology of yeast species invites characterization of genomic expression patterns to illuminate the details of cellular physiology. In addition to its value as an interesting organism, yeast maintains its role as an excellent model in which to characterize genomic expression programs. Microarray studies are quickly spreading to plant, animal, and microbial organisms that remain in the early stages of characterization. The extensive knowledge of yeast biology, as well as the relative ease with which yeast studies can be performed and controlled, facilitates interpretation of the genomic expression data. Importantly, existing information about yeast biology, including functional annotations for each gene, is captured and efficiently presented in databases such as the Saccharomyces Genome Database (SGD), the Munich Information Center Yeast Genome Database (MIPS), the Yeast and Pombe Protein Databases (YPD and PPD, respectively), and others. A number of databases also allow the exploration of published genomic expression studies, including the "Expression Connection" at SGD and the Microarray Global Viewer (yMGV) organized by Marc et al. Consulting these databases to retrieve known details about gene function and regulation vastly facilitates interpretation of the genomic expression data, allowing biological hypotheses to be formulated and tested. These hypotheses can be applied to other organisms that may execute genomic expression programs similar to those seen in yeast. Furthermore, as more genomic expression studies in multiple organisms emerge, large-scale data comparisons can be conducted, within and across organisms. Incorporating the results of yeast studies into such comparisons is certain to increase our understanding about the function, regulation, and evolution of genomic expression programs.

Carbocyanines↗
PubMed2001

Performance of mycological media in enumerating desiccated food spoilage yeasts: an interlaboratory study.

Dichloran 18% glycerol agar (DG18) was originally formulated to enumerate nonfastidious xerophilic moulds in foods containing rapidly growing Eurotium species. Some laboratories are now using DG18 as a general purpose medium for enumerating yeasts and moulds, although its performance in recovering yeasts from dry foods has not been evaluated. An interlaboratory study compared DG18 with dichloran rose bengal chloramphenicol agar (DRBC), plate count agar supplemented with chloramphenicol (PCAC), tryptone glucose yeast extract chloramphenicol agar (TGYC), acidified potato dextrose agar (APDA), and orange serum agar (OSA) for their suitability to enumerate 14 species of lyophilized yeasts. The coefficient of variation for among-laboratories repeatability within yeast was 1.39% and reproducibility of counts among laboratories was 7.1%. The order of performance of media for recovering yeasts was TGYC > PCAC = OSA > APDA > DRBC > DG 18. A second study was done to determine the combined effects of storage time and temperature on viability of yeasts and suitability of media for recovery. Higher viability was retained at -18 degrees C than at 5 degrees C or 25 degrees C for up to 42 weeks, although the difference in mean counts of yeasts stored at -18 degrees C and 25 degrees C was only 0.78 log10 cfu/ml of rehydrated suspension. TGYC was equal to PCAC and superior to the other four media in recovering yeasts stored at -18 degrees C, 5 degrees C, or 25 degrees C for up to 42 weeks. Results from both the interlaboratory study and the storage study support the use of TGYC for enumerating desiccated yeasts. DG18 is not recommended as a general purpose medium for recovering yeasts from a desiccated condition.

Agar↗
PubMed2003

Spoilage yeasts in the wine industry.

Yeasts play a central role in the spoilage of foods and beverages, mainly those with high acidity and reduced water activity (a(w)). A few species are capable of spoiling foods produced according to good manufacturing practices (GMPs). These can survive and grow under stress conditions where other microorganisms are not competitive. However, many of the aspects determining yeast spoilage have yet to be clarified. This critical review uses the wine industry as a case study where serious microbiological problems are caused by yeasts. First, the limitations of the available tools to assess the presence of spoilage yeasts in foods are discussed. Next, yeasts and factors promoting their colonisation in grapes and wines are discussed from the ecological perspective, demonstrating that a deeper knowledge of vineyard and winery ecosystems is essential to establish the origin of wine spoilage yeasts, their routes of contamination, critical points of yeast infection, and of course, their control. Further, zymological indicators are discussed as important tools to assess the microbiological quality of wines, although they are rarely used by the wine industry. The concepts of the susceptibility of wine to spoilage yeasts and wine stability are addressed based on scientific knowledge and industrial practices for monitoring yeast contamination. A discussion on acceptable levels of yeasts and microbiological criteria in the wine industry is supported by data obtained from wineries, wholesalers, and the scientific literature.Finally, future directions for applied research are proposed, involving collaboration between scientists and industry to improve the quality of wine and methods for monitoring the presence of yeast.

Culture Media↗
PubMed1998

Adhesive interactions between medically important yeasts and bacteria.

Yeasts are being increasingly identified as important organisms in human infections. Adhesive interactions between yeasts and bacteria may contribute to yeast retention at body sites. Methods for studying adhesive interactions between bacterial strains are well known, and range from simple macroscopic methods to flow chamber systems with complex image analysis capabilities. The adhesive interactions between bacteria and yeasts have been studied employing several of the methods originally developed for studying adhesive interactions between bacteria. However, in many of the methods employed the larger size of the yeasts as compared with bacteria results in strong sedimentation of the yeasts, often invalidating the method adapted. In addition, most methods are semi-quantitative and do not properly control mass transport. Consequently, adhesive interaction mechanisms between yeasts and bacteria identified hitherto, including lectin binding and protein-protein interactions, must be regarded with caution. Extensive physico-chemical characteristics of yeast cell surfaces are not available and a physico-chemical mechanism has not yet been put forth. A new method for quantifying adhesive interactions between yeasts and bacteria is proposed, based on the use of a parallel plate flow chamber, in which the influence of adhering bacteria upon the kinetics of yeast adhesion and aggregation of the adhering yeasts is quantitatively evaluated, under carefully controlled mass transport.

Bacteria↗
PubMed2006

Variable adhesion and diurnal population patterns of epiphytic yeasts on creeping bentgrass.

Irrigation and an in vitro agitation assay were used to determine the percentage of the epiphytic yeast community (Cryptococcus, Pseudozyma, Rhodotorula, and Sporobolomyces) adhering to the phylloplane of creeping bentgrass (Agrostis palustris (Huds.) Pers.). Colony-forming units (cfu) of total epiphytic yeast populations (adherent and nonadherent cells) and of adherent populations (cells not removed by agitation) were determined by leaf washing and dilution plating. In an in vitro assay, 40.0% and 57.1% of the yeast adhered to the leaves, whereas, in initial field trials the percentage of adherent yeasts ranged from 40.0% to 71.9% of the total population. Adherent yeast cfu on leaves in the morning were significantly lower on bentgrass (8.0 x 103 to 3.1 x 104 cfu.cm(-2)) compared with total yeast cfu (1.4 x 104 to 4.7 x 104 cfu.cm(-2)) on the nonirrigated control. No differences in yeast populations were observed between irrigated and nonirrigated plots 2 h after the 0900 treatments. Yeast populations followed a diurnal pattern, with larger cfu recovered from bentgrass leaves in the morning and significantly lower populations recovered in the afternoon. At 1400 the adherent yeast were 83.1%-100% of the total yeast population recovered from the leaves. The relative adhesiveness of the epiphytic yeast community on bentgrass leaves is dynamic with nonadherent cells making up a larger percentage of the population in the mornings than the afternoons.

Agrostis↗
PubMed2001

Effects of forage neutral detergent fiber and yeast culture on performance of cows during early lactation.

Sixty Holstein cows were assigned to two treatments at 21 d before calving and were group-fed a prepartum diet with or without yeast culture. After parturition, cows were individually fed one of five treatments for 140 d: 1) 21% forage neutral detergent fiber (NDF) without yeast culture, 2) 21% forage NDF with yeast culture, 3) 17% forage NDF without yeast culture, 4) 17% forage NDF with yeast culture, and 5) 25% forage NDF with yeast culture for 30 d and then switched to diet 4 for 110 d. Cows fed yeast culture prepartum were also fed yeast culture postpartum (60 g/d). A quadratic increase to 25, 21, and 17% forage NDF occurred during the first 30 d in milk (DIM) for dry matter intake, milk yield, and milk protein yield. No differences were observed for yeast culture or interaction of yeast culture and forage NDF for the first 30 DIM. Feeding 17 versus 21% forage NDF increased milk protein percentage and tended to increase dry matter intake as a percentage of body weight from 31 to 140 DIM. During this period, yeast culture tended to increase milk fat percentage and appeared to have positive effects on dry matter intake, milk yield, and milk fat yield when supplemented to diets with 21% forage NDF but not with 17% forage NDF. Feeding 17% forage NDF may be too low for the first 30 DIM but may improve animal performance after 30 DIM compared to 21% forage NDF.

Animals↗
PubMed2004

Enumeration of yeasts in dairy products: a comparison of immunological and genetic techniques.

Enzyme-linked immunosorbent assay (ELISA) and PCR techniques have been developed for the detection of spoilage yeast species in dairy products. Polyclonal antibodies against live yeast cells (AY) were raised in rabbits by inoculation of a mixture of 10 yeast species frequently associated with dairy products spoilage. AY antibodies were used for the development of two ELISA formats (indirect and double-antibody sandwich ELISA) for the detection of yeast species in milk and yogurt. A PCR assay was also developed for yeast detection in dairy products, using primers designed to amplify a conserved 250-base pair fragment of the 18S rRNA of the yeast species. The results obtained in this work show that ELISA techniques using polyclonal antibodies against viable yeast cells are of limited value for the detection and enumeration of spoilage yeast species in dairy products. On the contrary, PCR amplification of a conserved region of the 18S rRNA of the yeast species allows the homogeneous detection of all the yeast species tested and, combined with an overnight enrichment of samples, could be used for the detection of low levels of viable spoilage yeast species in dairy products.

Colony Count, Microbial↗
PubMed2006

Comparative analysis of regulatory transcription factors in Schizosaccharomyces pombe and budding yeasts.

Regulatory transcription factors (rTFs), which bind specific DNA sequences in the regulatory regions of genes and subsequently activate or repress transcription, play a central role in programming genomic expression. The number of rTFs in a species might therefore reflect its functional complexity. For simple organisms like yeast, a relatively small number of rTFs might be expected that is fairly constant between yeast species. We show that the budding yeast, Saccharomyces cerevisiae, contains 201 rTfs, which is one of the largest rTF numbers found in yeast species for which genome sequences are available. This is a much higher number than the 129 rTFs found in the fission yeast, Schizosaccharomyces pombe, which is currently the yeast with the lowest number of rTFs. Comparative analysis of several different budding yeast species shows that most of the 'extra' rTFs found in S. cerevisiae were probably acquired as a result of a whole genome duplication (WGD) event that occurred in an ancestor of a subset of budding yeast species. However, we also show that budding yeast species that have not been affected by the WGD contain a greater number of rTFs than S. pombe (mean = 145). Thus, two or more mechanisms have led to the 60% increase in rTFs in S. cerevisiae compared to S. pombe. This difference may correlate with a more extensive functional divergence in budding yeasts compared to fission yeasts. The relatively small number of rTFs in S. pombe make this organism an attractive model for global studies of mechanisms that programme gene expression.

Evolution, Molecular↗
PubMed1997

Establishment of a novel host, high-red yeast that stably expresses hamster NADPH-cytochrome P450 oxidoreductase: usefulness for examination of the function of mammalian cytochrome P450.

A novel strain of Saccharomyces cerevisiae useful for expression studies of mammalian microsomal cytochrome P450s was established and named High-red yeast. Hamster NADPH-cytochrome P450 oxidoreductase (P450 reductase) cDNA to be introduced into yeast was isolated from a hamster liver cDNA library. The cDNA was 2421 bp long and contained an entire coding region for 667 amino acids. The NH2-terminal amino acid sequence deduced from the hamster P450 reductase cDNA was identical with that of the enzyme purified from hamster livers except for deletion of the initial methionine. A delta-sequence derived from yeast retrotransposon Ty was cloned and used as a sequence for homologous recombination in a yeast genome. S. cerevisiae YPH500 was transformed with a multi-integration cassette containing the expression unit of the hamster P450 reductase and the delta-sequence. The transformant showing the highest activity of the P450 reductase was named High-red yeast. High-red yeast carried more than six copies of the multi-integration cassettes in a single chromosome and retained the multi-integration cassettes over a period of 100 generations under nonselective culture conditions, indicating that this yeast was a mitotically stable transformant. The microsomes prepared from High-red yeast had 20 times the P450 reductase activity of the microsomes prepared from the parental yeast. Due to the high activity of the hamster P450 reductase, the 7-ethoxycoumarin deethylase activity of mouse CYP1A1 expressed in High-red yeast was 250 times higher than the activity of mouse CYP1A1 expressed in the parental yeast.

Amino Acid Sequence↗
PubMed1993

In vitro susceptibility of mycelial and yeast forms of Penicillium marneffei to amphotericin B, fluconazole, 5-fluorocytosine and itraconazole.

The mycelial (25 degrees C) and yeast-like (37 degrees C) forms of Penicillium marneffei clinical and type strains were investigated for their in vitro susceptibility to amphotericin B (AmB), 5-fluorocytosine (5-FC), fluconazole (FLU) and itraconazole (ITZ), using Bacto antibiotic medium 3, yeast-nitrogen, Sabouraud's dextrose (pH 5.7) and high resolution (pH 7.1) broth media (1ml/tube), respectively. Results indicated that the minimal inhibitory and minimal fungicidal concentrations (MICs and MFCs) for the mycelial cultures of P. marneffei to AmB were in the range 0.78-1.56 and 0.78-3.125 micrograms/ml, respectively, as against 3.125-25 micrograms (MICs) for the yeast form cultures. The MFCs to AmB for the yeast form were one dilution higher. The MICs to FLU were generally lower for the yeast form (6.25-25 micrograms) than the mycelial form (25-50 micrograms/ml), whereas MFCs for the mycelial cultures were > 100 micrograms as compared to 6.25-100 micrograms for their yeast form. The MICs for the mycelial form to 5-FC ranged from < 0.195-0.39 microgram. Higher MICs (6.25 micrograms) were recorded for their yeast form. The MFCs to 5-FC for the yeast form were 25-100 micrograms/ml. The MICs for the mycelial form to ITZ ranged from < 0.195 to 3.125 micrograms/ml. Higher values (< 0.195-50 micrograms) were recorded for their yeast-like form. The MFCs to ITZ for mycelial and yeast forms ranged from < 0.195-0.39 and 25-100 micrograms/ml, respectively. Results indicate that P. marneffei's yeast form is more sensitive to FLU and ITZ (8 of 10 strains) while the mycelial form displayed greater susceptibility to AmB and 5-FC. The MICs for ITZ remained steady in SD medium, pH 5.7 to 7.1. However, some strains gave higher MIC values (0.39-1.56 micrograms/ml) when tested in the HR.

Amphotericin B↗
PubMed1993

Catabolism of benzene compounds by ascomycetous and basidiomycetous yeasts and yeastlike fungi. A literature review and an experimental approach.

A literature review is given on growth of yeasts on benzene compounds and on the catabolic pathways involved. Additionally, a yeast collection was screened for assimilation of phenol and 3-hydroxybenzoic acid. Fifteen ascomycetous and thirteen basidiomycetous yeast species were selected and were tested for growth on 84 benzene compounds. It appeared that 63 of these compounds supported growth of one or more yeast species. The black yeast Exophiala jeanselmei assimilated 54 of these compounds. The catechol branch of the 3-oxoadipate pathway and its hydroxyhydroquinone variant were involved in phenol and resorcinol catabolism of ascomycetes as well as of basidiomycetes. However, these two groups of yeasts showed characteristic differences in hydroxybenzoate catabolism. In the yeastlike fungus E. jeanselmei and in basidiomycetes of the genera Cryptococcus, Leucosporidium and Rhodotorula, the protocatechuate branch of the 3-oxoadipate pathway was induced by growth on 3- and 4-hydroxybenzoic acids. In three Trichosporon species and in all ascomycetous yeasts tested, 4-hydroxybenzoic acid was catabolyzed via protocatechuate and hydroxyhydroquinone. These yeasts were unable to cleave protocatechuate. 3-Hydroxybenzoic and 3-hydroxycinnamic acids were catabolized in ascomycetous yeasts via the gentisate pathway, but in basidiomycetes via protocatechuate. Incomplete oxidation of phenol, some chlorophenols, cresols and xylenols was observed in cultures of Candida parapsilosis growing on hydroquinone. Most compounds transformed by the growing culture were also converted by the phenol monooxygenase present in cell-free extracts of this yeast. They did not support growth. The relationship between the ability of ascomycetous yeasts to assimilate n-alkanes, amines and benzene compounds, and the presence of Coenzyme Q9 is discussed.

Ascomycota↗
PubMed2001

HO gene polymorphism in Saccharomyces industrial yeasts and application of novel HO genes to convert homothallism to heterothallism in combination with the mating-type detection cassette.

Southern blot analysis of industrial yeasts showed that all top-fermenting yeasts, distiller's yeasts and a proportion of wine yeasts tested in the present study produced a hybridization signal (approximately 7 kb), corresponding to a Saccharomyces cerevisiae-type HO gene (Sc-HO). It also showed that bottom-fermenting yeasts gave rise to 7-kb and 4-kb hybridization signals, corresponding to the Sc-HO gene and the lager yeast HO gene (Lg-HO), respectively. Two wine yeasts produced a 4-kb hybridization signal, corresponding to Lg-HO; and one wine yeast produced 2.5-kb and 1.5-kb hybridization bands, corresponding to a S. uvarum-type HO gene (Uv-HO). Partial nucleotide sequences of HO genes amplified from these wine yeasts perfectly matched those of Lg-HO and Uv-HO, respectively. HO disruption vectors were constructed by inserting a dominant selective marker PGK1p-neo and the mating-type detection cassette MFalpha1p-PHO5 within the Lg-HO or Uv-HO gene. From transformants carrying a single-disrupted ho gene, mating-competent progenies were easily obtained through meiosis. Moreover, mating-competent derivatives appearing at very low frequency could be obtained from a double-disrupted ho transformant without meiosis (even from a wine yeast lacking sporulation ability), because the sensitive phosphatase-staining method allowed detection of the Pho+ mating-competent derivatives from confluent colonies by the random spore method. Our study describes a rapid and convenient method for isolating mating-competent clones from industrial yeasts.

Amino Acid Sequence↗
PubMed1997

Complementation of temperature-sensitive topoisomerase II mutations in Saccharomyces cerevisiae by a human TOP2 beta construct allows the study of topoisomerase II beta inhibitors in yeast.

We show herein that human DNA topoisomerase II beta is functional in yeast. It can complement a yeast temperature-sensitive mutation in topoisomerase II. The effect on human topoisomerase II beta of a number of topoisomerase II inhibitors was analysed in a yeast in vivo system and compared with that of human topoisomerase II alpha and wild-type yeast topoisomerase II. A drug permeable yeast strain (JN394 top2-4) was used to analyse the in vivo effects of known anti-topoisomerase II agents on human topoisomerase II beta transformants. A parallel analysis on human topoisomerase II alpha transformants provides the first in vivo analysis of the responses of yeast bearing the individual isoforms to these drugs. The strain was analysed at 35 degrees C, a non-permissive temperature at which only plasmid-borne topoisomerase II is active. A shuttle vector with either human topoisomerase II beta, human topoisomerase II alpha or yeast topoisomerase II under the control of a GAL1 promoter was used. The key findings were that amsacrine produced comparable levels of cell killing with both alpha and beta, whilst etoposide, doxorubicin and mitoxantrone produced higher degrees of cell killing with alpha than with beta or yeast topoisomerase II. Merbarone had the greatest effect on the yeast strain bearing plasmid-borne yeast topoisomerase II. Suramin, quercetin and genistein showed little cell killing in this system. This yeast in vivo system provides a powerful way to analyse the effects of anti-topoisomerase II agents on transformants bearing the individual human isoforms. This system also provides a means of analysing putative drug-resistance mutations in human topoisomerase II beta or to select for drug-resistance mutations in human topoisomerase II beta.

Amsacrine↗
PubMed1985

In yeast, RAS proteins are controlling elements of adenylate cyclase.

S. cerevisiae strains containing RAS2val19, a RAS2 gene with a missense mutation analogous to one that activates the transforming potential of mammalian ras genes, have growth and biochemical properties strikingly similar to yeast strains carrying IAC or bcy1. Yeast strains carrying the IAC mutation have elevated levels of adenylate cyclase activity. bcy1 is a mutation that suppresses the lethality in adenylate cyclase deficient yeast. Yeast strains deficient in RAS function exhibit properties similar to adenylate cyclase deficient yeast. bcy1 suppresses lethality in ras1- ras2- yeast. Compared to wild-type yeast strains, intracellular cyclic AMP levels are significantly elevated in RAS2val19 strains, significantly depressed in ras2- strains, and virtually undetectable in ras1- ras2- bcy1 strains. Membranes from ras1- ras2- bcy1 yeast lack the GTP-stimulated adenylate cyclase activity present in membranes from wild-type cells, and membranes from RAS2val19 yeast strains have elevated levels of an apparently GTP-independent adenylate cyclase activity. Mixing membranes from ras1- ras2- yeast with membranes from adenylate cyclase deficient yeast reconstitutes a GTP-dependent adenylate cyclase.

Adenylyl Cyclases↗
PubMed2006

Shear-flow induced detachment of Saccharomyces cerevisiae from stainless steel: influence of yeast and solid surface properties.

The present study focused on the shear-induced detachment of Saccharomyces cerevisiae in adhesive contact with a 316L stainless steel surface using a shear stress flow chamber, with a view to determining the respective influence of the yeast surface properties and the support characteristics. The effect of cultivation of S. cerevisiae yeast cells on their subsequent detachment from the solid surface was particularly investigated. In order to elucidate the role of stainless steel, non-metallic supports were used as control, covering a broad range of surface properties such as surface free energy and roughness: polypropylene (hydrophobic), polystyrene (mildly hydrophobic, similar to stainless steel) and glass (hydrophilic). All materials were very smooth with respect to the size of yeast. First, experiments were carried out on two types of yeast cells, just rehydrated in saline solution, a biological model widely used in the literature. The influence of the ionic strength (1.5 and 150 mM NaCl) on glass and stainless steel was evaluated. Unlike on glass, no clear evidence was found for electrostatic repulsion with stainless steel since high adhesion was observed whatever the ionic strength. A lack of correlation in adhesion results was also obtained when considering the surface physico-chemical characteristics of type I (hydrophilic) and type II (hydrophobic) rehydrated cells and those of both polymers. It was postulated that unavoidable "sticky" compounds were present on the cell wall, which could not be completely removed during the successive washings of the rehydrated cell suspension before use. This could dramatically alter the yeast surface properties and modify the adhesion strength, thus clearly demonstrating the necessity to work with yeast coming from fresh cultures. Biologically active yeast cells were then used. Once cultured, type I- and type II-yeast cells were shown to exhibit the same hydrophilic properties. Regardless of the material used, for the same ionic strength (150 mM NaCl), yeast adhesion was drastically reduced compared to rehydrated yeast cells. Among all the materials tested, the specificity of 316L stainless steel was clearly established. Indeed, for glass and polymers, cell adhesion was substratum-dependent and driven by the balance between the Lifshitz-van der Waals and Lewis acid/base interactions. Despite nearly identical surface free energies for polystyrene and stainless steel, the metallic surface promoted a totally distinct behaviour which was characterized by a strong - although highly variable - yeast adhesion.

Cell Adhesion↗
PubMed1998

Evaluating the effect of soft lining materials on the growth of yeast.

STATEMENT OF PROBLEM: Soft lining materials continue to have a place in clinical removable prosthodontics. However, there is an increased probability of yeast colonization on soft lining materials. PURPOSE: This study (1) assessed a method of evaluating the effect of long-term soft lining materials on the growth of yeast and (2) investigated the effect five soft lining materials had on the growth of three species of yeast. MATERIAL AND METHODS: Coe Supersoft, Novus, and three experimental soft lining materials were investigated together with Candida albicans, Candida tropicalis, and Issatchenkia orientalis (formerly Candida krusei) yeasts. Strips of soft lining material incubated on blood agar plates were examined for inhibition of the growth of yeast. Soft lining materials soaked in sterile trypticase soya broth or water were inoculated with yeast and incubated. The change in colony forming units per milliliter from the initial load of yeast at 3 days was measured. Statistical analysis was performed with an independent paired Student t test. RESULTS: Inhibition of yeast growth occurred for two soft lining materials. Despite the presence of sufficient viable organisms, differences between the initial load of yeast and the 3-day results were mostly small, both for the test and control groups, suggesting that the material does not support the growth of the tested yeast during this period. CONCLUSIONS: The often described increased prevalence of yeast associated with soft lining materials in the oral environment is likely related to readily available nutrients in the mouth and the difficulty in maintaining and cleaning these materials.

Acrylic Resins↗