ABSTRACT The indiscriminate use of pesticides on grape crops is harmful for consumers´ healthin “in natura” consumption and in the ingestion of wine and grape juice. During winemaking, a rapid and ...efficient fermentation stage is critical to avoid proliferation of contaminating microorganisms and to guarantee the product´s quality. Polymerase chain reaction (PCR) has the advantage of detecting these contaminants in the early stages of fermentation. However,this enzymatic reaction may also be susceptible to specific problems, reducing its efficiency. Agricultural practices, such as fungicide treatments, may be a source of PCR inhibiting factors and may also interfere in the normal course of fermentation.The action of the pesticides captan and folpet on PCR and on yeast metabolism was evaluated, once these phthalimide compounds are widely employed in Brazilian vineyards. DNA amplification was only observed at 75 and 37.5 µg/mL of captan concentrations, whereas with folpet, amplification was observed only in the two lowest concentrations tested (42.2 and 21.1µg/mL).Besides the strong inhibition on Taq polymerase activity, phthalimides also inhibited yeast metabolism at all concentrations analyzed.Grape must containing captan and folpet residues could not be transformed into wine due to stuck fermentation caused by the inhibition of yeast metabolism. Non-compliance with the waiting period for phthalimide fungicides may result in financial liabilities to the viticulture sector.The use of yeasts with high fungicide sensitivity should be selected for must fermentation as a strategy for sustainable wine production and to assure that products comply with health and food safety standards.
The evolutionary relationship of indoleamine 2,3-dioxygenase (IDO) to some gastropod myoglobins suggests that IDO may undergo autoxidation in vivo such that one or more currently unidentified ...electron donors are required to maintain IDO heme iron in the active, ferrous state. To evaluate this hypothesis we have used yeast knockout mutants in combination with a recently developed yeast growth assay for IDO activity in vivo to demonstrate a role for cytochrome b5 and cytochrome b5 reductase in maintaining IDO activity in vivo.
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BFBNIB, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Saccharomyces cerevisiae
CEN.PK113-5D, a strain auxotrophic for uracil belonging to the CEN.PK family of the yeast
S. cerevisiae
, was cultured in aerated fed-batch reactor as such and once ...transformed to express human interleukin-1β (IL-1β), aiming at obtaining high cell densities and optimizing IL-1β production. Three different exponentially increasing glucose feeding profiles were tested, all of them “in theory” promoting respiratory metabolism to obtain high biomass/product yield. A non-structured non-segregated model was developed to describe the performance of
S. cerevisiae
CEN.PK113-5D during the fed-batch process and, in particular, its capability to metabolize simultaneously glucose and ethanol which derived from the precedent batch growth. Our study showed that the proliferative capacity of the yeast population declined along the fed-batch run, as shown by the exponentially decreasing specific growth rates on glucose. Further, a shift towards fermentative metabolism occurred. This shift took place earlier the higher was the feed rate and was more pronounced in the case of the recombinant strain. Determination of some physiological markers (acetate production, intracellular ROS accumulation, catalase activity and cell viability) showed that neither poor oxygenation nor oxidative stress was responsible for the decreased specific growth rate, nor for the shift to fermentative metabolism.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
► We demonstrate that HCV core protein (Core) impairs growth in budding yeast. ► We identify HSP90 inhibitors as compounds that restore the yeast growth. ► HSC90 can confer stability of the nascent ...but not the pre-existing Core in yeast. ► An organelle interaction domain of the Core is crucial for the growth retardation. ► The yeast system may be utilized to select compounds that can reduce Core stability.
Hepatitis C virus core protein (Core) contributes to HCV pathogenicity. Here, we demonstrate that Core impairs growth in budding yeast. We identify HSP90 inhibitors as compounds that reduce intracellular Core protein level and restore yeast growth. Our results suggest that HSC90 (Hsc82) may function in the protection of the nascent Core polypeptide against degradation in yeast and the C-terminal region of Core corresponding to the organelle-interaction domain was responsible for Hsc82-dependent stability. The yeast system may be utilized to select compounds that can direct the C-terminal region to reduce the stability of Core protein.
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BFBNIB, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Microbial pectinolytic strains are targeted as potential starter to control cocoa fermentation. This study analyses the ability of yeasts pectinolytic strains to grow under stress conditions. After ...an initial growth, a decline trend was observed in yeast growth cycle, during the cocoa fermentation. The 36 yeasts pectinolytic strains screened from 205 isolates showed tolerance to both alcoholic stress (10-12% alcohol) and thermic stress (up to 40 degrees Celsius) corresponding to surviving population round 75%. However, temperature-alcohol cross stress provokes full inhibition of strains that failed to grow at only 35 degrees Celsius under 8-10% alcohol. As acid stress, citric acid at 4% has the same effect as alcohol. In contrast, acetic acid and lactic acid respectively at 0.5% and 2% exerted individually, a higher pressure on yeast growth inhibiting up to 60% the fungal population. However, the viability of yeasts strains to a given concentration of lactic (1%) or acetic (0.25%) acid proved to be relatively stable with YS201 at increasing temperature up to 40 degrees Celsius.
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CEKLJ, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
The influence of different concentrations of two new fungicides (metiram and pyraclostrobin) on the fermentative activity of Saccharomyces cerevisiae yeast during winemaking process has been ...evaluated through in vitro assays. These fungicides were assayed as pure active compounds (single and in combination) and as a commercial formulation which contains both fungicides (55% metiram + 5% pyraclostrobin). The presence of pyraclostrobin pure standard in the culture medium, at the highest concentration evaluated (10 mg/L), increased the biomass and ethanol production rate. No effect on the alcoholic fermentation was observed for metiram pure standard due to its low solubility in the synthetic medium. However, a total inhibition of yeast growth was observed in presence of greater than or equal to 40 mg/L of the commercial formulation.Original Abstract: Se analizo, mediante ensayos in vitro, la influencia de diferentes concentraciones de dos nuevos fungicidas - metiram y piraclostrobin - sobre la actividad fermentativa de la levadura Saccharomyces cerevisiae en la elaboracion de vino. Estos fungicidas fueron evaluados como materias activas puras, tanto de forma individual como combinada, y como producto fitosanitario comercial formulado con ambas materias activas (55% metiram + 5% piraclostrobin). La presencia de piraclostrobin en el medio de cultivo a la concentracion mas alta evaluada (10 mg/L) provoco una mayor produccion de biomasa y etanol. El metiram, debido a su baja solubilidad en el medio sintetico, no mostro ningun efecto sobre la fermentacion alcoholica. Por otro lado, concentraciones greater than or equal to 40 mg/L de la formulacion comercial provocaron una inhibicion total del crecimiento de la levadura.
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IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Cytoplasmic male sterility (CMS) has often been associated with abnormal mitochondrial open reading frames (ORF), orfH79 is a mitochondria chimeric gene being responsible for the CMS trait in ...Honglian (HL) rice. Weakly expressed ORFH79 strongly inhibits the growth of yeast cells. In addition, the content of reactive oxygen species (ROS) in the transformants that expressed ORFH79 was increased by 31%, and ATP was decreased by 41% compared with the control. These results showed ORFH79 peptide is toxic to yeast cells.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Rapid microscale toxicity tests make it possible to screen large numbers of compounds and greatly simplify toxicity identification evaluation and other effect directed chemical analyses of effluents ...or environmental samples. Tests using Vibrio fischeri (such as Microtox®) detect toxicants that cause non-specific narcosis, but are insensitive to other important classes of contaminants. The microbial assay for risk assessment (MARA) is a 24 h multi-species test that seeks to address this problem by using a battery of ten bacteria and a fungus. But there has been little independent evaluation of this test, and there is no published information on its sensitivity to pesticides. Here, we assess the performance of MARA using a range of toxicants including reference chemicals, fungicides and environmental samples. Mean MARA microbial toxic concentrations and IC₂₀s (20% Inhibitory concentrations) indicate the toxicant concentrations affecting the more sensitive micro-organisms, while the mean IC₅₀ (50% Inhibitory concentration) was found to be the concentration that was toxic to most MARA species. For the two fungicides tested, the yeast (Pichia anomalia) was the most sensitive of the ten MARA species, and was more sensitive than the nine other yeasts tested. The test may be particularly valuable for work with fungicides. Mean MARA IC₅₀s were comparable to values for nine other yeast species and the lowest individual IC₅₀s for each toxicant were comparable to reported IC₅₀s for Daphnia magna, Selenastrum capricornutum and Microtox® bioassays. MARA organisms exhibited more variable sensitivities, with the most sensitive organism being different for different samples, enhancing the likelihood of toxicity detection and giving a toxicity “fingerprint” that may help identify toxicants. The test, therefore, has great potential and would be valuable for ecotoxicological testing of pollutants.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
In the present work two separation techniques, namely the gravitational field-flow fractionation (GrFFF) and the reversed-flow gas chromatography (RFGC), are proposed for the distinction of the ...growth phases of
Saccharomyces cerevisiae (AXAZ-1) yeast cycle at different temperatures (30
°C, 25
°C, 20
°C, and 15
°C) and pH (2.0, 3.0, 4.0 and 5.0) values. During the fermentation processes, differences observed in the peak profiles, obtained by GrFFF, can be related with the unlike cell growth. The distinction of the phases of AXAZ-1 cell cycle with the GrFFF, was also confirmed with the RFGC technique, which presented similar fermentation time periods for the alcoholic fermentation phases. Simultaneously, the reaction rate constant for each phase of the fermentation process and the activation energies were determined with the aid of the RFGC technique. Finally, the application of both the GrFFF and the RFGC techniques, in combination with high-performance liquid chromatography, allowed us to find the ideal experimental conditions (temperature and pH) for the alcoholic fermentation by AXAZ-1. The results indicate that
S. cerevisiae cells performed better at 30
°C, whereas at lower temperatures decreases in the fermentation rate and in the number of viable cells were observed. Moreover, the pH of the medium (pH 5.0) resulted in higher fermentation rates and ethanol productivities.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
Mao wine is produced from mao (Antidesma thwaitesianum) and is a local beverage promoting Sakon Nakhon province, Thailand. Understanding the fermentation process is necessary in order to enhance the ...quality of mao wine products. This study aims to examine the kinetics of mao wine fermentation using 3 commercial types of Saccharomyces cerevisiae yeasts, namely Lalvin 71B, Lalvin RC212 and Lalvin K1-V1116, with an initial pH of 3.4, a sweetness of 22 % Brix and a yeast concentration of 0.4 g L–1 in controlled conditions at 25 °C for 10 days. The results revealed that the kinetic models, including logistic, Luedeking-Piret and modified Luedeking-Piret equations, are appropriate for analyzing the kinetics of yeast growth, alcohol production and reducing sugar, respectively. Lalvin K1-V1116 yeast yielded the highest capacity for alcohol production (~113 g L–1), sugar-to-alcohol conversion (45 % conversion), and product efficiency (88 %), indicating its suitability for mao wine production. Furthermore, the investigation of the alpha-amylase inhibitory activity revealed that the IC50 values of mao wine samples were 2.88 - 4.78 mg mL–1, which were roughly 410 - 680 times greater than those of acarbose. These findings revealed the low efficacy of mao wine in inhibiting alpha-amylase activity. Nevertheless, a decrease in the level of inhibition may result in a decreased likelihood of side effects or undesirable reactions. This treatment is especially beneficial for people who have medication sensitivities. Additional research is required in order to substantiate its applicability in the management of type 2 diabetes. HIGHLIGHTS Mao wines were produced by 3 commercial cerevisiae yeast strains Lalvin K1-V1116 yeast exhibits the highest capacity in converting sugar to alcohol Lalvin K1-V1116 is the most suitable yeast for making mao wine The chosen kinetic models are appropriate for kinetic studies of fermentation process The efficacy of Mao wine in inhibiting the activity of alpha-amylase is quite low GRAPHICAL ABSTRACT
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
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