Saccharomyces cerevisiae

High pressure homogenization (HPH) was tested for inducing autolysis in a commercial strain of Saccharomyces bayanus for winemaking. The effects on cell viability, the release of soluble proteins, glucidic colloids and amino acids in wine-like medium and the volatile composition of the autolysates were investigated after processing, in comparison with thermolysis. HPH seemed a promising technique for inducing autolysis of wine yeasts. One pass at 150MPa was the best operating conditions. Soluble colloids, proteins and free amino acids were similar after HPH and thermolysis, but the former gave a more interesting volatile composition after processing, with higher concentrations of ethyl esters (fruity odors) and lower fatty acids (potential off-flavors). This might allow different winemaking applications for HPH, such as the production of yeast derivatives for wine ageing. In the conditions tested, HPH did not allow the complete inactivation of yeast cells; the treatment shall be optimized before winemaking use.

The DNA damage response triggered by bacterial cytolethal distending toxins (CDTs) is associated with activation of the actin-regulating protein RhoA and phosphorylation of the downstream-regulated mitogen-activated protein kinase (MAPK) p38, which promotes the survival of intoxicated (i.e. cells exposed to a bacterial toxin) cells. To identify the effectors of this CDT-induced survival response, we screened a library of 4492 Saccharomyces cerevisiae mutants that carry deletions in nonessential genes for reduced growth following inducible expression of CdtB. We identified 78 genes whose deletion confers hypersensitivity to toxin. Bioinformatics analysis revealed that DNA repair and endocytosis were the two most overrepresented signaling pathways. Among the human orthologs present in our data set, FEN1 and TSG101 regulate DNA repair and endocytosis, respectively, and also share common interacting partners with RhoA. We further demonstrate that FEN1, but not TSG101, regulates cell sur...

The entire pathway of palmitate synthesis from malonyl-CoA in mammals is catalyzed by a single, homodimeric, multifunctional protein, the fatty acid synthase. Each subunit contains three N-terminal domains, the β-ketoacyl synthase, malonyl/acetyl transferase and dehydrase separated by a structural core from four C-terminal domains, the enoyl reductase, β-ketoacyl reductase, acyl carrier protein and thiosterase. The kinetics and specificities of the substrate loading reaction catalyzed by the malonyl/acetyl transferase, the condensation reaction catalyzed by β-ketoacyl synthase and chain-terminating reaction catalyzed by the thioesterase ensure that intermediates do not leak off the enzyme, saturated chains exclusively are elongated and palmitate is released as the major product. Only in the fatty acid synthase dimer do the subunits adopt conformations that facilitate productive coupling of the individual reactions for fatty acid synthesis at the two acyl carrier protein centers. Introduction of a double tagging and dual affinity chromatographic procedure has permitted the engineering and isolation of heterodimeric fatty acid synthases carrying different mutations on each subunit. Characterization of these heterodimers, by activity assays and chemical cross-linking, has been exploited to map the functional topology of the protein. The results reveal that the two acyl carrier protein domains engage in substrate loading and condensation reactions catalyzed by the malonyl/acetyl transferase and β-ketoacyl synthase domains of either subunit. In contrast, the reactions involved in processing of the β-carbon atom, following each chain elongation step, together with the release of palmitate, are catalyzed by the cooperation of the acyl carrier protein with catalytic domains of the same subunit. These findings suggest a revised model for the fatty acid synthase in which the two polypeptides are oriented such that head-to-tail contacts are formed both between and within subunits.

Yeast cells immobilized by entrapment in an insolubilized gelatin gel have been investigated with regard to their pattern of growth and respiratory activity. Mass-transfer resistance offered by the matrix and growth of the entrapped cells determine a gradient of nutrients throughout the gel which is responsible for both a lower specific growth rate of immobilized cells with respect to that of free ones, and a heterogeneous biomass distribution, with progressively increased cellular density from the inside to the outside of the matrix. The spatial organization of the growing cells leads to the formation of a biofilm perfectly adherent to the surface of the matrix. Measurements of the oxygen-uptake rate of the immobilized cells, after having been released from the matrix by trypsin digestion, and confocal microscopy of cells stained with Rhodamine 123 demonstrate the occurrence of a gradient of respiratory activity throughout the immobilized culture.

Lignocellulosic ethanol has been considered as an alternative transportation fuel. Utilization of hemicellulosic fraction in lignocelluloses is crucial in economical production of lignocellulosic ethanol. However, this fraction has not efficiently been utilized by traditional yeast Saccharomyces cerevisiae. Genetically modified S. cerevisiae, which can utilize xylose, has several limitations including low ethanol yield, redox imbalance, and undesired metabolite formation similar to native xylose utilizing yeasts. Besides, xylose uptake is a major issue, where sugar transport system plays an important role. These genetically modified and wild-type yeast strains have further been engineered for improved xylose uptake. Various techniques have been employed to facilitate the xylose transportation in these strains. The present review is focused on the sugar transport machineries, mechanisms of xylose transport, limitations and how to deal with xylose transport for xylose assimilation in ...

Identification of components present in biological complexes requires their purification to near homogeneity. Methods of purification vary from protein to protein, making it impossible to design a general purification strategy valid for all cases. We have devel-oped the tandem affinity purification (TAP) method as a tool that allows rapid purification under native conditions of complexes, even when expressed at their natural level. Prior knowledge of complex composition or function is not required. The TAP method requires fusion of the TAP tag, either N- or C-terminally, to the target protein of interest. Starting from a relatively small number of cells, active macromolecular complexes can be isolated and used for multiple applications. Variations of the method to specifi-cally purify complexes containing two given components or to subtract undesired complexes can easily be implemented. The TAP method was initially developed in yeast but can be successfully

Six different blends of lactic acid bacterial cultures (Lactobacillus plantarum or Pediococcus cerevisiae) combined with Saccharomyces cerevisiae were used as co-culture for preparing sourdoughs. These prefermented sourdoughs were applied for producing pan breads. Acidification power, leavening ability, and microbial counts were conducted at different sour fermentation intervals. The highest acidification power in sourdoughs were recorded when 2% wd/wd (dry weight

In the present study, the chemical composition and antioxidant potential of an essential oil of ginger rhizomes from Ecuador was elucidated. The analysis of the essential oil by GC/FID/MS resulted in identification of 71 compounds, of which the main are citral (geranial 10.5% and neral 9.1%), α-zingiberene (17.4%), camphene (7.8%), α-farnesene (6.8%) and β-sesquiphellandrene (6.7%). The in vitro antioxidant activity of the essential oil expressed by IC50 in descending order is: hydroxyl radical (OH*) scavenging (0.0065 μg/mL) > chelating capacity (0.822 μg/mL) > 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical cation (ABTS*+) scavenging (3.94 μg/mL) > xanthine oxidase inhibition (138.0 μg/mL) > oxygen radical (O2*) scavenging (404.0 μg/mL) > 2,2- diphenyl-1-picrylhydrazyl radical (DPPH*) scavenging (675 μg/mL). Lipid peroxidation inhibition of the essential oil was less efficient than butylhydroxytoluol (BHT) in both stages, i.e. hydroperoxide and malondia...

In this work, a brewer’s yeast strain was used to remove heavy metals from a synthetic effluent. The solid–liquid separation process was carried out using the flocculation ability of the strain. The yeast strain was able to sediment in the presence of Cu2+, Ni2+, Zn2+, Cd2+ and Cr3+, which evidences that the flocculation can be used as a cheap and natural separation process for an enlarged range of industrial effluents. For a biomass concentration higher than 0.5 g/l, more than 95% of the cells were settled after 5 min; this fact shows that the auto-aggregation of yeast biomass is a rapid and efficient separation process. Cells inactivated at 45 °C maintain the sedimentation characteristics, while cells inactivated at 80 °C lose partially (40%) the flocculation.The passage of metal-loaded effluent through a series of sequential batches allowed, after the second batch, the reduction of the Ni2+concentration in solution for values below the legal limit of discharge of wastewater in natural waters (2 mg/l); this procedure corresponds to a removal of 91%. A subsequent batch had a marginal effect on Ni2+ removal (96%).Together, the results obtained suggest that the use of brewing flocculent biomass looks a promising alternative in the bioremediation of metal-loaded industrial effluents since the removal of the heavy metals and cell separation are simultaneously achieved.

Natural flavonoids are currently receiving much attention because of their estrogenic and antiestrogenic properties. Six isoflavones (isoprunetin, isoprunetin 7-O-β-d-glucopyranoside, isoprunetin 4′,7-di-O-β-d-glucopyranoside, genistein, genistein 7-O-β-d-glucopyranoside, daidzein), four flavones (luteolin, luteolin 7-O-β-d-glucopyranoside, luteolin 4′-O-β-d-glucopyranoside, licoflavone C), isolated from Genista morisii and G. ephedroides (two Leguminosae plants of the Mediterranean area) together with two structurally related pterocarpans, bitucarpin A and erybraedyn C, isolated from Bituminaria bituminosa (Leguminosae), were tested for the antagonist activity by a yeast based estrogen receptor assay (Saccharomyces cerevisiae RMY326 ER-ERE). Most compounds inhibited the estradiol-induced transcriptional activity in a concentration dependent manner. In particular, for the flavone luteolin 77% inhibition of the induced β-galactosidase activity was observed. Interestingly, licoflavone C exhibited a dose-dependent antagonistic activity at concentrations up to 10−4 m, but stimulated β-galactosidase expression at higher concentrations resulting in a U-shaped-like dose-response curve. Copyright © 2007 John Wiley & Sons, Ltd.