Viability Of Saccharomyces Cerevisiae Research Articles

Protective properties of the edible halophyte Carpobrotus edulis (L.) N.E.Br. towards neoformed food contaminants-related oxidative stress and genotoxicity

Carpobrotus edulis (L.) N.E.Br. is an edible halophyte native to South Africa, where it has been traditionally consumed as food, revealing its nutritional value, and used in the treatment of human diseases probably due to the accumulation of bioactive compounds such as polyphenols. These compounds have potential to display valuable biological activities that could bring several benefits to the human being, including the mitigation of toxicity induced by food contaminants. Therefore, this study investigated biological activities with health-promoting potential of a C. edulis ethanol extract (CEE), such as antioxidant, antigenotoxic, and antimutagenic. CEE was shown by LC-DAD-ESI/MSn to be composed of 16 polyphenols. The extract exhibited antioxidant properties by protecting Saccharomyces cerevisiae viability and improving the redox state of HepG2 cells under oxidative stress. Antigenotoxicity of CEE against H2O2-induced oxidative damage was detected in HepG2 and Caco-2 cells by comet assay and subsequently observed in the dominant deletion assay in S. cerevisiae. Antigenotoxicity and antimutagenicity of CEE towards the neoformed contaminant benzo[a]pyrene was found in HepG2 and Caco-2 cells by comet assay and in Salmonella typhimurium using the Ames test, respectively. The disclosed antigenotoxic and antimutagenic properties of C. edulis further advance its potential for application in food and nutraceutical industries for health benefits.

Viability of Saccharomyces cerevisiae during baking of bread dough by flow cytometry

Flow cytometry analysis (FCA) is increasingly used to obtain rapid results comparison to common colony-forming units plating method (CFU). Tools are needed for microbiological analysis for solid matrix such as food. Here, we report a fast and robust FCA using double staining with LDS751/DiBAC4(3) to analyze yeast viability in bread dough during baking.

THE EFFECT OF ARGON ON THE YEAST UNDER CAVITATION

Introduction. The process of water purification from contaminants, in particular microbiological, is still relevant. One of the promising and effective physical methods of water disinfection is the use of cavitation, which is characterized by numerous positive results in water purification technology.Aim. To investigate the effect of cavitation and inert argon on the viability of Saccharomyces cerevisiae in water. To determine the effectiveness of argon during cavitation treatment of the water system.Methods. Ultrasonic generator UZDN-2T was used for cavitation treatment of yeast-containing water at frequency of 22 kHz, power 35 W, T = 298 ± 1 K, P = 0.1 MPa, the total duration of the process was 2 hours. A suspension of yeast cells in sterile water taken from a natural water body was used for research. Inert gas (argon) was bubbled into the water throughout the process with simultaneous action of cavitation. The results of calculating the number of the cells before and after the experiments are expressed in colony-forming units.The main results of the study. Treatment of water with high content of yeast with simultaneous action of inert gas and ultrasonic cavitation was carried out. The studied microobjects were Saccharomyces cerevisiae yeast with an initial content of 2.07×104 cells per unit volume of water. It is shown that after 30 min of Ar/US action the number of the microorganisms decreased 2.6 times relative to its initial number. The efficiency of the combined action of argon and cavitation was 99.5% (NMend = 100 CFU/cm3) in the end of the process. Conclusions. Cavitation action in an argon atmosphere on the water purification from yeast was shown. The proportions of dead cells which allowed to describe the process of water disinfection were calculated and active water purification under conditions of argon supply with high yeast content in 1 cm3 of aqueous medium was noted. Almost complete purification of water after two hours of treatment was achieved.Key words: water; cleansing; drіzhі; argon; cavitation

APPLICATION OF REFRIGERATED AND FROZEN SORGHUM MALT SLURRIES IN THE PRESERVATION OF STARTER CULTURES FOR OBUSHERA FROM UGANDA

Industrial production of traditional fermented beverages is limited by lack of quality commercial starter cultures. Saccharomyces cerevisiae MNC21Y and Lactobacillus plantarum MNC21 can be used to ferment cereal beverages such as Obushera . These cultures are unavailable as commercial starters due to lack of appropriate propagating and distributing procedures. The purpose of this study was to evaluate the use of refrigerated and frozen sorghum slurries as carrier media for the starters. Starters were propagated in sorghum slurries (30°C for 24 h) and stored at 5°C and -18°C for 90 days. Viability of Saccharomyces cerevisiae and Lactobacillus plantarum was determined by cell counts after surface plating and pour plating, respectively. Fermentation ability was determined by inoculating sorghum slurries with starters and monitoring pH, acidity and flavor development . Viability was higher for starters stored at 5°C (S. cerevisiae : 6 log cfu.g -1 and Lb. plantarum : 7-9 log cfu.g -1 during 90 days) than those at -18°C (S. cerevisiae : 2 cfu.g -1 and Lb. plantarum : 4 log cfu.g -1 after 30 days). Refrigerated starters acidified Obushera (pH £ 4.5) faster (10-20 h) than frozen ones (18-24 h). Refrigerated or frozen S. cerevisiae + Lb. plantarum starters in sorghum malt slurries can remain viable for at least one or three months, respectively and produce Obushera with characteristic flavors.

Radio frequency-induced superoxide accumulation affected the growth and viability of Saccharomyces cerevisiae.

With the development of the electric technologies, the biological effects of electromagnetic fields (EMF) were widely studied. However, the results remain controversial and the biophysical mechanisms are still unknown. To our knowledge, little studies pay attention to the radio frequency (RF) of 2.6-5MHz. In the present study, we investigated the effect of these radio frequencies on the growth and cell viability of Saccharomyces cerevisiae at very low power density below 0.1mT. The result appeared to be time-dependent. The growth of the yeast cells was obviously affected by the RF-EMF with a 43.5% increase when exposed for 30h, and the growth-promoting effect decreased along with the radiation time and eventually turned to an inhibiting effect retarding growth by 20.7% at 89h. The cell viability was improved to 70.1% at 8h and reduced by 33.5% at 28h. The superoxide accumulated in exposed cells as radiation time increased which may lead to the inhibition of viability and growth of the cells. However, the efficient frequency, power density, and exposure dosage await further investigation. Nevertheless, the wave band studied in this research is effective to produce biological effect, and therefore, it may provide an optional new radio frequency which is valuable for the development and utilization in therapy technique and medical use.

Validation of methylene blue viability staining with the emerging pathogen Candida auris

Methylene blue viability staining has been traditionally used to assess viability of Saccharomyces cerevisiae in brewing and wine making. Here, this method was tested and validated with the emerging fungal pathogen Candida auris to determine if this species would also deferentially stain, which could provide utility in assessing microbial control and disinfectant efficacy.

Expression of Metazoan Annexins in Yeast Provides Protection Against Deleterious Effects of the Biofuel Isobutanol

The ability of microorganisms to produce biofuels by fermentation is adversely affected by the perturbing effects of the hydrophobic biofuel on plasma membrane structure. It is demonstrated here that heterologous expression of metazoan, calcium-dependent, membrane-binding proteins of the annexin class can reduce deleterious effects of isobutanol on Saccharomyces cerevisiae viability and complex membrane functions. Therefore, expression of annexins in industrial strains of yeast or bacteria may prove beneficial in biofuel production.

Implication of a Novel Function of Sar1 in the Nucleus.

The coat protein complex II (COPII) generates transport carriers that deliver newly synthesized proteins from the endoplasmic reticulum (ER) to the Golgi apparatus. The small GTPase Sar1 is a well-known regulator of the assembly of the COPII coat. In the present study, we demonstrate that, besides its well-established role in ER-to-Golgi trafficking, the nuclear localization of Sar1 is essential for the viability of Saccharomyces cerevisiae. Inhibition of either the nuclear entry or retention of Sar1 leads to a severe growth defect. Additionally, in vivo deletion of Sar1, by using conditional genetic depletion, further demonstrates that the loss of nuclear localization of Sar1 results in an abnormal nuclear envelope shape. Our findings highlighted a possible novel role of Sar1 within the nucleus, which may relate to the proper formation of the nuclear envelope.Key words: Sar1, COPII, small GTPase, nuclear envelope, membrane traffic.

Monitoring of Saccharomyces cerevisiae viability by non-Faradaic impedance spectroscopy using interdigitated screen-printed platinum electrodes

Abstract Interdigitated, screen-printed platinum microelectrodes are used for non-Faradaic impedance spectroscopy detection of viable Saccharomyces cerevisiae . The time dependence of the complex impedance of both living and heat-inactivated yeast cells immersed in plain medium or immobilized in agar is measured between 0.01 Hz and 1 MHz for 300 min. To understand the processes probed by the impedance measurement, an equivalent electrical circuit, containing a constant phase element, a charge transfer resistance and a suspension resistance, is established and analyzed. For both heat-inactivated and living yeast cells in plain medium, the suspension resistance increases over time caused by sedimentation of the yeast cells into the stray field of the electrodes. The measured increase is found to be larger for living cells than for heat-inactivated ones. The time dependence of the double-layer capacitance is rather dominated by the metabolic activity of living cells, but a possible impact of cell sedimentation cannot be completely neglected. In experiments carried out with cells immobilized in agar, cell sedimentation is suppressed, and thus, the time evolution of the double-layer capacitance is solely caused by metabolic active cells. Therefore, the time-dependent change of the double-layer capacitance can be used to directly monitor the viability of Saccharomyces cerevisiae in situ , since inactivated cells do not contribute to the signal generation in this case.

Effects of the dose and viability of Saccharomyces cerevisiae. 2. Ruminal fermentation, performance of lactating dairy cows, and correlations between ruminal bacteria abundance and performance measures

This study examined effects of the dose and viability of supplemented Saccharomyces cerevisiae yeast strain YE1496 on ruminal fermentation and performance of lactating dairy cows. A second objective was to examine correlations between ruminal bacteria abundance and performance measures. Four ruminally cannulated lactating cows (284 ± 18 days in milk) were assigned randomly to 1 of 4 treatment sequences in a 4 × 4 Latin square experimental design using four 21-d experimental periods. Cows were fed a nonacidotic total mixed ration comprising 22.5% starch (minimum ruminal pH >5.8), 41.7% corn silage, 7.60% wet brewers grain, and 50.7% concentrate on a dry matter (DM) basis. The diet was supplemented with no yeast (control), a low (5.7 × 107 cfu/d; LLY) or high (6.0 × 108 cfu/d; HLY) dose of live yeast, or a high dose of killed yeast (6.0 × 108 cfu/d; killed by heating at 80°C for 1.5 h; HDY). Milk production and composition were measured twice daily from d 11 to 21 of each period, and rumen fluid samples were collected on d 21. In vivo digestibility was measured using chromic oxide as a marker. Pearson correlation analysis was used to assess whether animal performance parameters were correlated with relative abundance (RA) of ruminal bacteria. Supplemental LLY increased yields (kg/d) of milk (29.6 vs. 31.7) and milk protein (0.95 vs. 1.03), tended to increase milk fat yield (1.10 vs. 1.17) and ruminal acetate:propionate ratio (1.92 vs. 2.21), and increased in vivo apparent digestibility (%) of DM (64.5 vs. 69.1), neutral detergent fiber (NDF; 45.0 vs. 54.5), and ADF (53.1 vs. 60.9) compared with the control. Feeding HLY had no effects on milk yield compared with the control (30.0 vs. 29.6 kg/d). Feeding HDY tended to increase in vivo digestibility (%) of NDF (45.0 vs. 50.7), ADF (53.1 vs. 57.7), and the ruminal concentration of lactate (0.78 vs. 2.82 mM) but did not affect milk yield compared with the control. Dry matter and NDF digestibility correlated negatively with RA of unclassified Lachnospiraceae in both solid (r = -0.50 and -0.52, respectively) and liquid (r = -0.56 and -0.57, respectively) fractions, whereas milk yield correlated positively with RA of Lachnospiraceae [Ruminococcus] (an incompletely classified genus; r = 0.43) in the solid ruminal fraction. Supplemental LLY, HLY, or HDY increased or tended to increase DM, NDF, and ADF digestibility, but only LLY increased yields of milk, milk fat, and milk protein.

Effects of the dose and viability of Saccharomyces cerevisiae. 1. Diversity of ruminal microbes as analyzed by Illumina MiSeq sequencing and quantitative PCR

This study was conducted to examine effects of the dose and viability of supplemental Saccharomyces cerevisiae on the ruminal fermentation and bacteria population and the performance of lactating dairy cows. Four ruminally cannulated lactating cows averaging 284±18d in milk were assigned to 4 treatments arranged in a 4×4 Latin square design with four 21-d periods. Cows were fed a total mixed ration containing 41.7% corn silage, 12.1% brewer’s grains, and 46.2% concentrate on a dry matter basis. The diet was supplemented with no yeast (control) or with a low dose of live yeast (5.7×107 cfu/cow per day; LLY), a high dose of live yeast (6.0×108 cfu/cow per day; HLY), or a high dose of killed yeast (6.0×108 cfu/cow per day; HDY). Microbial diversity was examined by high-throughput Illumina MiSeq sequencing (Illumina Inc., San Diego, CA) of the V4 region of the 16S rRNA gene. The relative abundance of select ruminal bacteria was also quantified by quantitative PCR (qPCR). Adding LLY to the diet increased the relative abundance of some ruminal cellulolytic bacteria (Ruminococcus and Fibrobacter succinogenes) and amylolytic bacteria (Ruminobacter, Bifidobacterium, and Selenomonas ruminantium). Adding live instead of killed yeast increased the relative abundance of Ruminococcus and F. succinogenes; adding HDY increased the relative abundance of Ruminobacter, Bifidobacterium, Streptococcus bovis, and Selenomonas ruminantium. The most dominant (≥1% of total sequences) bacteria that responded to LLY addition whose functions are among the least understood in relation to the mode of action of yeast include Paraprevotellaceae, CF231, Treponema, and Lachnospiraceae. Future studies should aim to speciate, culture, and examine the function of these bacteria to better understand their roles in the mode of action of yeast. A relatively precise relationship was detected between the relative abundance of F. succinogenes (R2=0.67) from qPCR and MiSeq sequencing, but weak relationships were detected for Megasphaera elsdenii, Ruminococcus flavefaciens, and S. ruminantium (R2≤0.19).

Pyrolysis Byproducts as Feedstocks for Fermentative Biofuel Production: An Evaluation of Inhibitory Compounds through a Synthetic Aqueous Phase

In this work, the viability of using aqueous-phase sugars derived from the pyrolysis of lignocellulosic biomass was analyzed using high-throughput screening in microtiter plates. To standardize results, a synthetic aqueous phase of pyrolytic bio-oil was constructed based on typical constituents and composition ranges and then used to determine the fermentation viability of Saccharomyces cerevisiae. The effects of inhibitory compounds in pyrolytic bio-oil were assessed by fitting measured growth kinetics to the model of Baranyi and Roberts (Int. J. Food Microbiol. 1994, 23, 277), specifically on the fitted growth rates, initial microorganism adaptation, and maximum biomass densities. It was found that even a dilution to approximately 10% of the hypothetical inhibitor concentration in aqueous bio-oil was significantly inhibitory to growth, although the presence of additional sugars was able to moderate this impact slightly. The high-throughput screening used in this work allowed for the rapid measurement of...

Viability of Saccharomyces cerevisiae incorporated within silica and polysaccharide hosts monitored via time-resolved fluorescence

The viability of Saccharomyces cerevisiae in biocompatible polymers under different growth conditions and studied using time-resolved fluorescence techniques is presented. Two fluorophores, the viscosity sensitive probe 4-(4-(dimethylamino)styryl)-N-methyl-pyridiniumiodine (DASPMI) and the yeast viability stain 2-chloro-4-(2,3-dihydro-3-methyl-(benzo-1,3-thiazol-2-yl)-methylidene)-1-phenylquinolinium iodide (FUN-1) are used to elucidate information on the incorporated yeast cell viability. Variations in cell viscosity, which are indicative of the cell state, were obtained using DASPMI. Prior to observing FUN-1 in yeast cells using fluorescence lifetime imaging, its photophysics in solution and heterogeneous media were investigated. Time-resolved emission spectra were measured and analysed to associate lifetimes to the spectral emission. Preliminary results show that monitoring the fluorescence lifetime of FUN-1 may give a useful insight into cellular metabolism. The results indicate that both fluorophores may be used to monitor the entrapped yeast cell viability, which is important for in vitro studies and applications, such as that in the biofuel industry, where Saccharomyces cerevisiae are required to remain active in high ethanol environments.

Crambescidin-816 Acts as a Fungicidal with More Potency than Crambescidin-800 and -830, Inducing Cell Cycle Arrest, Increased Cell Size and Apoptosis in Saccharomyces cerevisiae

In this paper, we show the effect of crambescidin-816, -800, and -830 on Saccharomyces cerevisiae viability. We determined that, of the three molecules tested, crambescidin-816 was the most potent. Based on this result, we continued by determining the effect of crambescidin-816 on the cell cycle of this yeast. The compound induced cell cycle arrest in G2/M followed by an increase in cell DNA content and size. When the type of cell death was analyzed, we observed that crambescidin-816 induced apoptosis. The antifungal effect indicates that crambescidins, and mostly crambescidin-816, could serve as a lead compound to fight fungal infections.

The consequences of Lactobacillus vini and Dekkera bruxellensis as contaminants of the sugarcane-based ethanol fermentation

This work describes the effects of the presence of the yeast Dekkera bruxellensis and the bacterium Lactobacillus vini on the industrial production of ethanol from sugarcane fermentation. Both contaminants were quantified in industrial samples, and their presence was correlated to a decrease in ethanol concentration and accumulation of sugar. Then, laboratory mixed-cell fermentations were carried out to evaluate the effects of these presumed contaminants on the viability of Saccharomyces cerevisiae and the overall ethanol yield. The results showed that high residual sugar seemed the most significant factor arising from the presence of D. bruxellensis in the industrial process when compared to pure S. cerevisiae cultures. Moreover, when L. vini was added to S. cerevisiae cultures it did not appear to affect the yeast cells by any kind of antagonistic effect under stable fermentations. In addition, when L. vini was added to D. bruxellensis cultures, it showed signs of being able to stimulate the fermentative activity of the yeast cells in a way that led to an increase in the ethanol yield.

Survival rate of wine-related yeasts during alcoholic fermentation assessed by direct live/dead staining combined with fluorescence in situ hybridization

Real-time detection of microorganisms involved in complex microbial process, such as wine fermentations, and evaluation of their physiological state is crucial to predict whether or not those microbial species will be able to impact the final product. In the present work we used a direct live/dead staining (LDS) procedure combined with fluorescence in situ hybridization (FISH) to simultaneously assess the identity and viability of Saccharomyces cerevisiae (Sc) and Hanseniaspora guilliermondii (Hg) during fermentations performed with single and mixed cultures. The population evolution of both yeasts was determined by plating and by LDS combined with species-specific FISH-probes labeled with Fluorescein. Since the FISH method involves the permeabilization of the cell membrane prior to hybridization and that it may influence the free diffusion of PI in and out of the cells, we optimized the concentration of this dye (0.5μg of PI per 106 cells) for minimal diffusion (less than 2%). Fluorescent cells were enumerated by hemocytometry and flow cytometry. Results showed that the survival rate of Sc during mixed cultures was high throughout the entire process (60% of viable cells at the 9th day), while Hg began to die off at the 2nd day, exhibited 98% of dead cells at the 3rd day (45g/l of ethanol) and became completely unculturable at the 4th day. However, under single culture fermentation the survival rate and culturability of Hg decreased at a much slower pace, exhibiting at the 7th day (67g/l of ethanol) 8.7×104CFU/ml and 85% of dead cells. Thus, our work demonstrated that the LDS–FISH method is able to simultaneously assess the viability and identity of these wine-related yeast species during alcoholic fermentation in a fast and reliable way. In order to validate PI-staining as a viability marker during alcoholic fermentation, we evaluated the effect of ethanol on the membrane permeability of Sc and Hg cells, as well as their capacity to recover membrane integrity after being exposed to different levels of ethanol (1%, 6%, 10%, 12% v/v). Results showed that while Sc cells were able to recover membrane integrity after ethanol exposure, Hg cells were not. However, under alcoholic fermentation Sc cells didn't recover membrane integrity after the mid-term (4–5days) of alcoholic fermentation.

A microplate-based method for real-time monitoring of Saccharomyces cerevisiae viability

Measuring the viability of the cells after chemical or physical stresses is a routine method in the determination of phenotypes in yeast. Numerous analytical techniques have been applied to the measurement of yeast cell viability, but few of them are suitable for real-time monitoring. Here we propose a microplate-based method for real-time monitoring of Saccharomyces cerevisiae cell viability. This method is also suitable for high-throughput measurement.

The influence of carboxymethylcellulose, xanthan and guar-gum addition in bread dough before freezing on metabolism and viability of Saccharomyces cerevisiae

Doughs were prepared with different concentrations of carboxymethylcellulose, xanthan, and guar-gum (0.1, 0.2 and 0.3% in doughs), freezed at -20?C and analyzed after 0, 7, 15 and 30 days. Pure Saccharomyces cerevisiae culture was isolated from dough and was cultivated under optimal conditions during 24 hrs to determine the following parameters: specific growth rate, fermentative activities and cytochromes contents in intact cells with the aim of determining the respiration intensity. During freezing of dough for 30 days, the percentage of living cells from dough surface was 53.11% and from the middle 54.95%. Carboxymethylcellulose in concentration of 0.3 and 0.5% increased number of survived cells on the surface to 70.64, and 70.28% and in the middle to 74.79, and 76.54%, respectively. Guar-gum increased number of survived cells only in concentration of 0.1% on the surface to 70.17% and in the middle of the dough to 75.26%. The mean specific growth rate decreased by approximately 10% during 30 days of storage at -20?C. Content of cytochromes in intact cells decreased in all samples during freezing.

Expression of Human Protein Phosphatase-1 in Saccharomyces cerevisiae Highlights the Role of Phosphatase Isoforms in Regulating Eukaryotic Functions

Human (PP1) isoforms, PP1alpha, PP1beta, PP1gamma1, and PP1gamma2, differ in primary sequences at N and C termini that potentially bind cellular regulators and define their physiological functions. The GLC7 gene encodes the PP1 catalytic subunit with >80% sequence identity to human PP1 and is essential for viability of Saccharomyces cerevisiae. In yeast, Glc7p regulates glycogen and protein synthesis, actin cytoskeleton, gene expression, and cell division. We substituted human PP1 for Glc7p in yeast to investigate the ability of individual isoforms to catalyze Glc7p functions. S. cerevisiae expressing human PP1 isoforms were viable. PP1alpha-expressing yeast grew more rapidly than strains expressing other isoforms. On the other hand, PP1alpha-expressing yeast accumulated less glycogen than PP1beta-or PP1gamma1-expressing yeast. Yeast expressing human PP1 were indistinguishable from WT yeast in glucose derepression. However, unlike WT yeast, strains expressing human PP1 failed to sporulate. Analysis of chimeric PP1alpha/beta subunits highlighted a critical role for their unique N termini in defining PP1alpha and PP1beta functions in yeast. Biochemical studies established that the differing association of PP1 isoforms with the yeast glycogen-targeting subunit, Gac1p, accounted for their differences in glycogen synthesis. In contrast to human PP1 expressed in Escherichia coli, enzymes expressed in yeast displayed in vitro biochemical properties closely resembling PP1 from mammalian tissues. Thus, PP1 expression in yeast should facilitate future structure-function studies of this protein serine/threonine phosphatase.

Yeast cell inactivation related to local heating induced by low-intensity electric fields with long-duration pulses

The effects of electric field (EF) treatments on Saccharomyces cerevisiae viability were investigated using a PG200 electroporator (Hoefer Scientific Instrument, San Fransisco, CA, USA) with specific attention to induced thermal effects on cell death. Lethal electric fields (1.5 kV cm − 1 for 5 s) were shown to cause heat variations in the cell suspension medium (water + glycerol), while corresponding classical thermal treatments at equivalent temperatures had no effect on the cells viability. Variations of the electrical conductivity of the intra- and extracellular matrix caused by ions and solutes transfer across the membrane were shown to be involved in the observed heating. The results permitted to build a theoretical model for the temperature variations induced by electric fields. Using this model and the electrical conductivity of the different media, a plausible explanation of the cell death induced by low-intensity electric fields with long-duration pulses has been proposed. Indeed, cell mortality could in part be caused by direct and indirect effects of electric fields. Direct effects are related to well known electromechanical phenomena, whereas indirect effects are related to secondary thermal stress caused by plasma membrane thermoporation. This thermoporation was attributed to electrical conductivity variations and the corresponding intracellular heating.