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PubMed1997

The effect of dietary active dry yeast supplement on performance of sows during gestation-lactation and their pigs.

Thirty crossbred sows and their pigs were evaluated through two parities to determine any reproductive or growth performance effects of an active dry yeast supplement added to corn-soybean meal diets. Sow reproductive performance from d 93 of gestation through d 21 of lactation and sow milk composition were evaluated. Pig growth performance was measured from birth to 28 d after weaning. Active dry yeast was added at 0, 1, or .2% of the sow gestation diet, 0, .15, or .3% of the sow lactation diet, 0, .2, or .4% of the pig prestarter diet, 1 wk before and 1 wk after weaning, and 0, .125, or .25% during the last 3 wk in the nursery. The yeast source consisted of a concentrate of live yeast cells of the Saccharomyces cerevisiae strain containing more than 15 x 10(9) live cells/g. Sow body weight at d 93 of gestation, at farrowing, and at d 21 of lactation did not differ (P > .10) among treatment groups. Milk from sows fed active dry yeast contained higher amounts of total solids (P < .05), crude protein (P < .10), and gamma globulin (P < .06) than milk from sows fed the control diet. Sow feed intake during lactation was not affected (P > .10) by treatment, nor were there differences in litter size at birth, litter birth weight, or litter weight at d 21 after farrowing. Active dry yeast supplementation to the sow and pig diets resulted in improved postweaning pig daily gain (P < .05) and gain-to-feed ratio (P < .05) but did not affect (P > .10) feed intake. Based on these data, active dry yeast supplement during late gestation, lactation, and before and after weaning does not alter litter weight at birth or weaning but does increase gamma globulin content of sow's milk and improves postweaning rate and efficiency of weight gain of pigs.

Animals↗
PubMed1987

Conventional and rapid methods for yeast identification.

Identification of yeasts depends on data obtained from morphological features and physiological characteristics. This article reviews the commonly used conventional methods for the identification of yeast to set the stage for a critical review of rapid systems for yeast identification. Comparative analyses of percentage agreement of API 20C, Uni-Yeast Tek, Minitek, Mycotube, Micro-Drop, Auxodisk, Iatron, Autobac 1, AMS, Abbott MS-2, and Abbott Quantum 11 against the conventional method and, in some cases, against each other are tabulated. Gas liquid chromatography method, enzyme method, MUG, and the Fung's mini-system are also mentioned as possible rapid methods for yeast identification. The purpose of this review is to update the rapidly expanding information concerning diagnostic kits and systems for yeast identification and analyze some of the advantages, disadvantages and potential uses of rapid systems of yeast identification.

Mycology↗
PubMed2004

Feed efficiency of mid-lactation dairy cows fed yeast culture during summer.

Thirty-eight Holstein cows (26 multiparous and 12 primiparous), that averaged 105 d postpartum at the start of the experiment, were used to evaluate the feeding of yeast culture (60 g/cow daily of Diamond V XP) on production efficiency during hot summer weather. From early June until early September and after a 2-wk covariate period, cows were fed a control diet without or with 60 g of yeast culture/cow daily for 12 wk. Weekly daytime high temperatures in the free-stall barn during the 12-wk period averaged 33 degrees C (28 to 39 degrees C). Total mixed diets on a dry matter (DM) basis consisted of corn silage (28%), alfalfa hay (21%), and a concentrate mix (51%) without or with the yeast culture added to the total mixed ration at the time of feeding. Milk production (34.9 and 35.4 kg/d, for control and yeast culture treatment, respectively), 4% fat-corrected milk (31.2 and 32.0 kg/d), energy-corrected milk (ECM; 33.4 and 34.2 kg/d), and DM intake (23.1 and 22.1 kg/d) were similar for cows fed control and yeast culture diets. Percentages of milk fat (3.34 and 3.41) and true protein (2.85 and 2.87) were similar for both diets. Feed efficiency defined as kilogram of ECM/kilogram of DM intake was improved by 7% for cows fed the yeast culture. Body weights and body condition scores were similar for both groups. The results suggest that the yeast culture can improve feed efficiency of heat stressed dairy cows in midlactation.

Animal Feed↗
PubMed2005

Molecular epidemiological study on pre-X region of hepatitis B virus and identification of hepatocyte proteins interacting with whole-X protein by yeast two-hybrid.

AIM: To identify the pre-X region in hepatitis B virus (HBV) genome and to study the relationship between the genotype and the pre-X region. To investigate the biological function of whole-X (pre-X plus X) protein, we performed yeast two-hybrid to screen proteins in liver interacting with whole-X protein. METHODS: The pre-X region of HBV was amplified by polymerase chain reaction (PCR) method, and was cloned to pGEM Teasy vector. After the target region was sequenced, Vector 8.0 software was used to analyze the sequences. The whole-X bait plasmid was constructed by using yeast two-hybrid system 3. Yeast strain AH109 was transformed. After expression of the whole-X protein in AH109 yeast strains was proved, yeast two-hybrid screening was performed by mating AH109 with Y187 containing liver cDNA library plasmid. The mated yeast was plated on quadruple dropout medium and assayed for alpha-gal activity. The interaction between whole-X protein and the protein obtained from positive colonies was further confirmed by repeating yeast two-hybrid. After extracting and sequencing of plasmid from blue colonies, we carried out analysis by bioinformatics. RESULTS: After sequencing, 27 of 45 clones (60%) were found encoding the pre-X peptide. Eighteen of twenty-seven clones (66.7%) of pre-X coding sequences were found from genotype C. Five positive colonies that interacted with whole-X protein were obtained and sequenced; namely, fetuin B, UDP glycosyltransferase 1 family-polypeptide A9, mannose-P-dolichol utilization defect 1, fibrinogen-B beta polypeptide, transmembrane 4 superfamily member 4-CD81 (TM4SF4). CONCLUSION: The pre-X gene exists in HBV genome. Genes of proteins interacting with whole-X protein in hepatocytes were successfully cloned. These results brought some new clues for studying the biological functions of whole-X protein.

Base Sequence↗
PubMed1998

Yeast communities associated with sugarcane in Campos, Rio de Janeiro, Brazil.

Yeast communities associated with sugarcane leaves, stems and rhizosphere during different phases of plant development were studied near Campos, in Rio de Janeiro, Brazil. Atmospheric temperature, soil granulometry and pH, and sugar cane juice degree Brix and pH were determined. Yeast communities associated with sugarcane were obtained after cellular extraction by shaking, blending and shaking plus sonication, and cultured on Yeast Nitrogen Base Agar plus glucose (0.5%) and Yeast Extract-Malt Extract Agar. No significant differences in yeast counts were found among the cellular extraction treatments and culture media. 230 yeast cultures were identified according to standard methods, and distinct yeast communities were found for each substrate studied. The prevalent species isolated from sugarcane were Cryptococcus laurentii, Cryptococcus albidus, Rhodotorula mucilaginosa and Debaryomyces hansenii.

Brazil↗
PubMed1975

[The enzymatic hydrolysis of cell wall applied to yeasts classification (author's transl)].

The rigid structure of yeast cell wall may be a taxonomic criterion. Using the Helix pomatia juice (with mercaptoethylamine hydrochloride) as a polyenzymatic system, we have always obtained protoplasts from ascomycetous yeasts and never from basidiomycetous yeasts tested. Then, we have applied this enzymatic analysis to the Fungi imperfecti yeasts and, according to this test, separated genus looking like basidiomycetous behaviour: Trichosporon, Pityrosporum, Rhodotorula, Cryptococcus and non fermentative Candida (group II). Oppositely, we have put together the Fungi imperfecti yeasts looking like ascomycetous behaviour: Kloeckera, Trigonopsis, Brettanomyces and fermentative Candida (group I). With the second lytic system made of mercaptoethylamine-HCl and beta-(1-3)-D-glucanase, we have selected ascomycetous yeasts giving protoplasts: Saccharomycoidea with genus Saccharomyces, Kluyveromyces, Pichia, Hansenula, Citeromyces, Debaryomyces; we have joined Saccharomycodes and Hanseniaspora to this second group. The other ascomycetous yeasts studied did not give protoplasts with the definite lytic system made of purified beta-(1-3)-D-glucanase and thiol. These results seem to demonstrate that the structure of cell wall can act as an important taxonomic criterion.

Ascomycota↗
PubMed2003

[Screening the hepatitis B virus PreS1 associated protein by the yeast two-hybrid system].

OBJECTIVE: To screen the hepatitis B virus PreS1 associated protein from normal human liver cDNA library by the yeast two-hybrid system and explore the role of PreS1 protein in the infection of hepatitis B virus (HBV). METHODS: PCR was preformed to amplify the PreS1 gene containing EcoRI and PstI from HBV positive serum, and the production was inserted into plasmid pAS2-1 after digesting with the former two restricted endonuclease, then the bait vector pAS2-1-PreS1 was verified by auto-sequencing assay. The PreS1-BD fusion protein expressed in the yeast cells was confirmed by western blot, after pAS2-1-PreS1 was transfected into the yeast cell AH109. Yeast cells co-transfected with pAS2-1-PreS1 and the normal human liver cDNA library grew in selective SC/-trp-leu-his-ade2 medium, and the second screening was performed with LacZ report gene. Furthermore, segregation analysis and mating experiment were done to eliminate the false positive, then the true positive clones were submitted for PCR and sequencing. The results were submitted to the BLAST notebook of World Wide Wed Site NCBI to seek homologous sequence. RESULTS: Bait vector pAS2-1-PreS1 included the anticipated fragment of PreS1 gene. Western blot showed that pAS2-1-PreS1 could correctly express PreS1-BD fusion protein in the yeast cells. After yeast cells co-transfected with pAS2-1-PreS1 and the normal human liver cDNA library, 97 colonies grew in the selective SC/-trp-leu-his-ade2 medium, only one clone was positive and showed high homology with Homo sapiens nascent-polypeptide-associated complex alpha polypeptide. CONCLUSION: Bait vector pAS2-1-PreS1 is successfully constructed, and nascent-polypeptide-associated complex alpha polypeptide protein expressed in hepatocyte can interact with PreS1 by the yeast two-hybrid system.

Base Sequence↗
PubMed2002

Cloning and expression of the preS1 gene of hepatitis B virus in yeast cells.

OBJECTIVE: To investigate the complex functions of HBV preS1 protein, we constructed HBV preS1 gene expression vector and expressed it in yeast cells. METHODS: Polymerase chain reaction (PCR) was performed to amplify the gene of HBV preS1 from the plasmid pCP10 containing the whole DNA fragment of HBV ayw subtype as template and the PCR product was cloned into the pGEM-T vector for sequencing. After being identified, the HBV preS1 gene was cut from the pGEM-T vector by EcoR I and Pst I restriction enzymes, and cloned into yeast expressive plasmid pGBKT7 to construct pGBKT7-preS1 recombinant expressive plasmid. This plasmid was transformed into yeast cell AH109 and expressed in it. The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Western blotting. RESULTS: The HBV preS1 gene was amplified successfully and identified by DNA sequencing. The PCR products were coincided completely with the reported sequence. The digested fragments were cloned into the pGBKT7 vector and transformed into yeast cell AH109. The results of SDS-PAGE and Western blotting assay showed: (1) The HBV preS1 protein was expressed and existed in yeast cells; (2) The molecular weight of the expression product was about 30 000 D. CONCLUSION: The HBV preS1 gene was successfully cloned and expressed in yeast cells.

Blotting, Western↗
PubMedDate not supplied

Speciation and bioavailability of selenium in yeast-based intervention agents used in cancer chemoprevention studies.

This study investigated the speciation and bioavailability of selenium in yeast-based intervention agents from multiple manufacturers from several time points. Sources of selenized yeast included Nutrition 21 (San Diego, CA), which supplied the Nutritional Prevention of Cancer (NPC) Trial from 1981-1996; Cypress Systems (Fresno, CA; 1997-1999); and Pharma Nord (Vejle, Denmark; 1999-2000), which supplied the Prevention of Cancer by Intervention by Selenium (PRECISE) Trial pilot studies. The low-molecular-selenium species were liberated from the samples by proteolytic hydrolysis followed by separation by ion exchange liquid chromatography and detection by inductively coupled plasma-mass spectrometry. The results for the NPC tablets showed that selenomethionine, together with 3 unidentified selenium compounds, were predominant in the sample hydrolysates. The relative amounts of the 4 selenium species varied (p < 0.05) among several of the 7 tablet batches used during the course of the NPC Trial. In comparison, 5 batches of more recently produced selenized yeasts, which were used as a source of selenium in the PRECISE and other trials, contained less of the unknown compounds and more selenomethionine at 54-60% of the total selenium in the yeasts. One batch of yeast, however (from 1985), which originated from the same producer as the yeast used in the NPC tablets, contained only 27% of selenium in the sample as selenomethionine. Human subjects receiving 200 microg selenium/day in the UK PRECISE Pilot Trial showed a higher concentration (p < 0.01) and higher increase from baseline in plasma selenium than did the same dosage used in the NPC Trial. Differences in intake, speciation, or bioavailability of selenium from the yeast-based supplements in the population groups studied may explain this. Furthermore, the selenium concentration in whole blood from the Danish PRECISE Pilot Trial was higher (p < 0.001) than that obtained with synthetic L-selenomethionine in a comparable group of Danes, both groups having been treated with 300 microg selenium/day.

Anticarcinogenic Agents↗
PubMedDate not supplied

[Interrelationships between yeast fungi and collembolans in soil].

The possibility of feeding on green and newly fallen leaves of the small-leaved lime Tilia cordata was studied for the collembolans Protaphorura armata and Vertagopus pseudocinereus. Young leaves grown under sterile conditions and almost free of yeast fungi were established to be toxic to the collembolan V. pseudocinereus: feeding on them led to the death of the animals. Leaves grown under natural conditions were nontoxic: when used by the collembolans as feed, they provided for collembolan growth and fecundity. Feeding preferences of the collembolans in relation to the yeasts attributed to different ecomorphs-epiphytes, litter saprophytes, pedobionts, and saccharobionts-were studied. Of the 24 yeast strains isolated from plant green parts, litter, and soil and assigned to eight species, no strain was revealed that was not used by the collembolans. However, certain yeast strains were preferable for the collembolans. The population of the V. pseudocinereus collembolans feeding on the yeast Rhodotorula glutinis (nss 31-4) exceeded that grown on Cryptococcus terricola (2044) 1.5-fold. Thus, the collembolans have feeding preferences in relation to yeast fungi, as was shown earlier for mycelial micromycetes. The possible mechanisms of the feeding preferences of the collembolans in relation to yeasts are discussed.

Animal Nutritional Physiological Phenomena↗
PubMed1986

Selenium yeast.

Baker's yeast is able to assimilate carbon, nitrogen, phosphorus and sulphur sources together with a great number of minerals and trace elements into a palatable, nutritious product. The metabolism of yeast is precisely controlled during the production growth phase and thus it is possible to determine the composition of the product by controlling the raw materials. Because of existing deficiencies in the availability of certain trace elements, mainly selenium, in Finnish diets, we started testing the possibilities for enriching yeast with this essential trace element about five years ago. We have succeeded in developing a special yeast product with a selenium concentration of 500 mg/kg dry matter. Selenium was expected, because of its structural similarity to sulphur, to replace sulphur in the biosynthetic reactions of the yeast cell. We have recently studied the incorporation and distribution of selenium in yeast with radioactive selenium (75Se). Analysis of the protein fraction of selenium yeast has shown that selenium is present in all the major soluble proteins. Selenomethionine was identified as the major selenium-containing compound in the protein fraction as well as in the whole cell.

Amino Acids↗
PubMed1961

Purity control of the production of baker's yeast employing a fluorescent antiserum.

A simple staining procedure for the rapid detection of wild yeasts contaminating baker's yeast during the course of industrial production is described. Fluorescein-labeled, specific antiserum against Saccharomyces cerevisiae is applied to smears of baker's yeast which are then examined by ultraviolet microscopy. Optimal results are obtained with the combined phase contrast and fluorescence which makes the S. cerevisiae appear green, whereas cells of wild yeasts are visible in bright red counterstain. With this method, wild yeasts could be identified in baker's yeast at a dilution of 1:10,000.

Immune Sera↗
PubMed2003

Phagocytosis and peroxidase release by seabream (Sparus aurata L.) leucocytes in response to yeast cells.

A flow cytometric method was adapted to evaluate phagocytosis by gilthead seabream leucocytes after incubation with yeast cells (Saccharomyces cerevisiae). Head-kidney leucocytes were incubated in vitro for different times in different proportions with heat-killed fluorescein isothiocyanate (FITC)-labeled yeast cells to study the kinetics of phagocytosis. Attached and internalized yeast cells were differentiated by quenching FITC-labeled S. cerevisiae with trypan blue dye. Only internalized cells kept their FITC fluorescence after quenching. Monocyte-macrophages and acidophilic granulocytes showed phagocytic activity, as demonstrated by transmission electron microscopy (TEM). From the ultrastructural features of the phagocytic process, it was observed that cytoplasmic granule membranes fused with the phagocyte membrane at the point where the yeast cell was attached to the phagocyte surface. This observation led us to adapt a colorimetric method to study peroxidase (myeloperoxidase and eosinophil peroxidase) release, since both are considered to be markers of the degranulation that occurs in seabream head-kidney leucocytes in response to yeast cells. Head-kidney leucocytes were incubated with calcium ionophore (CaI), phorbol myristate acetate (PMA), or yeast cells for different periods of time (0-30 min) to study the kinetics of peroxidase release. The results obtained indicate that CaI and yeast cells, but not PMA, stimulate the degranulation (by about 44.51% and 21.04%, respectively, at 30 min) of seabream head-kidney leucocytes.

Animals↗
PubMed2001

Effects of 50 Hz magnetic field on cell cycle kinetics and the colony forming ability of budding yeast exposed to ultraviolet radiation.

To investigate the effects of extremely low frequency magnetic fields on ultraviolet radiation (UV) exposed budding yeast, haploid yeast (Saccharomyces cerevisiae) cells of the strain SEy2101a were exposed to 50 Hz sine wave magnetic field (MF) of 120 microT with simultaneous exposure to UV radiation. Most of the UV energy was in the UVB range (280-320 nm). The biologically weighted (CIE action spectrum) dose level for the UV radiation was 175 J/m2. We examined whether 50 Hz MF affected the ability of UV irradiated yeast cells to form colonies (Colony Forming Units, CFUs). In addition, the effect of coexposure on cell cycle kinetics was investigated. Although the significant effect of MF on the cell cycle phases of UV exposed yeast cells was seen only at one time point, the overall results showed that MF exposure may influence the cell cycle kinetics at the first cycle after UV irradiation. The effect of our particular MF exposure on the colony forming ability of the UV irradiated yeast cells was statistically significant 420 min after UV irradiation. Moreover, at 240, 360, and 420 min after UV irradiation, there were fewer CFUs in every experiment in (UV+MF) exposed populations than in only UV exposed yeast populations. These results could indicate that MF exposure in conjunction with UV may have some effects on yeast cell survival or growth.

Cell Cycle↗
PubMed2006

Yeast as a drug discovery platform in Huntington's and Parkinson's diseases.

The high degree of conservation of cellular and molecular processes between the budding yeast Saccharomyces cerevisiae and higher eukaryotes have made it a valuable system for numerous studies of the basic mechanisms behind devastating illnesses such as cancer, infectious disease, and neurodegenerative disorders. Several studies in yeast have already contributed to our basic understanding of cellular dysfunction in both Huntington's and Parkinson's disease. Functional genomics approaches currently being undertaken in yeast may lead to novel insights into the genes and pathways that modulate neuronal cell dysfunction and death in these diseases. In addition, the budding yeast constitutes a valuable system for identification of new drug targets, both via target-based and non-target-based drug screening. Importantly, yeast can be used as a cellular platform to analyze the cellular effects of candidate compounds, which is critical for the development of effective therapeutics. While the molecular mechanisms that underlie neurodegeneration will ultimately have to be tested in neuronal and animal models, there are several distinct advantages to using simple model organisms to elucidate fundamental aspects of protein aggregation, amyloid toxicity, and cellular dysfunction. Here, we review recent studies that have shown that amyloid formation by disease-causing proteins and many of the resulting cellular deficits can be faithfully recapitulated in yeast. In addition, we discuss new yeast-based techniques for screening candidate therapeutic compounds for Huntington's and Parkinson's diseases.

Amyloid↗
PubMed2006

Hydrocortisone made in yeast: metabolic engineering turns a unicellular microorganism into a drug-synthesizing factory.

Inspired by the successful work of converting Saccharomyces cerevisiae into an microorganism capable of synthesizing hydrocortisone, a 27-carbon molecule, from ethanol, a 2-carbon molecule, this review provides an overview of the potential of yeast as a recombinant organism in the 21st century. Yeast has been used by man for more than 6,000 years, and is still paving the way to new discoveries. It was the first eukaryotic organism to be sequenced, in 1996, and the first to produce hydrocortisone in 2003. In addition, extensive genome-wide analyses have been performed with yeast. In this review, we discuss the pros and cons of using yeast to produce small therapeutic molecules. It is obvious that S. cerevisiae has a cutting edge advantage of being a well-known organism and time will tell if yeast "biohydrocortisone" is a unique example or the beginning of a long list of yeast bioproducts. Other organisms, such as plants and bacteria, are competing with yeast. Bacteria produce a wealth of marketed molecules and plants are capable of producing extremely complex molecules with an unbeatable yield. However, S. cerevisiae offers a unique mix of the simplicity of a recombinant organism combined with the complexity of a eukaryote.

Biotechnology↗
PubMed2004

Physicochemical surface properties of brewing yeast influencing their immobilization onto spent grains in a continuous reactor.

Immobilization of brewing yeast onto a cellulose-based carrier obtained from spent grains, a brewing byproduct, by acid/base treatment has been studied in a continuously operating bubble-column reactor. The aim of this work was to study the mechanisms of brewing yeast immobilization onto spent grain particles through the information on physicochemical surface properties of brewing yeast and spent grain particles. Three mechanisms of brewing yeast immobilization onto spent grains carrier were proposed: cell-carrier adhesion, cell-cell attachment, and cell adsorption (accumulation) inside natural shelters (carrier's surface roughness). The possibility of stable cell-carrier adhesion regarding the free energy of interaction was proved and the relative importance of long-range forces (Derjaguin-Landau-Verwey-Overbeek theory) and interfacial free energies was discussed. As for the cell-cell attachment leading to a multilayer yeast immobilization, a physicochemical interaction through localized hydrophobic regions on cell surface was hypothesized. However, neither flocculation nor chain formation mechanism can be excluded so far. The adsorption of brewing yeast inside sufficiently large crevices (pores) was documented with photomicrographs. A positive effect of higher dilution rate and increased hydrophobicity of base-treated spent grains on the yeast immobilization rate has also been found.

Cell Adhesion↗
PubMed2001

3-Nitrocoumarin is an efficient inhibitor of budding yeast phospholipase-C.

3-Nitrocoumarin is described in the literature as a specific inhibitor of mammalian phospholipase-C and here we studied the effect of 3-nitrocoumarin on budding yeast phosphatidylinositol-specific phospholipase-C and its effect on yeast growth. 3-Nitrocoumarin is a powerful inhibitor in vitro of the yeast Plc1 protein with an IC(50) of 57 nM and it is also an inhibitor of yeast growth in minimal media at comparable concentrations. Moreover at the same concentration it inhibits the glucose-induced PI-turnover. Since the effects of 3-nitrocoumarin on yeast growth are superimposable on the growth phenotype caused by PLC1 gene deletion we can conclude that 3-nitrocoumarin is a specific and selective inhibitor of yeast phospholipase-C. In addition we show that 3-nitrocoumarin was also an effective inhibitor of the pathogenic yeast Candida albicans.

Candida albicans↗