Tolerant Yeast Research Articles

FLO Genes Family and Transcription Factor MIG1 Regulate Saccharomyces cerevisiae Biofilm Formation During Immobilized Fermentation.

Saccharomyces cerevisiae immobilization is commonly used for efficient ethanol fuel production in industry due to the relatively higher ethanol stress resistance of S. cerevisiae in biofilms relative to planktonic cells. The mechanisms of biofilm formation and stress resistance, however, remain ambiguous. By analyzing biofilm and planktonic cell transcriptomes, this study observed that MIG1 (encoding a transcription factor) expression in cells increases during the biofilm formation process. To identify the role of MIG1 in yeast biofilm formation and the ethanol resistance of these cells, MIG1 was deleted and complemented in S. cerevisiae 1308. Results showed the MIG1 deletion mutant strain demonstrated weaker biofilm formation ability both on fibers and plastic than the wild-type and these could be restored by expressing MIG1 in deletion mutant. To verify the ability of MIG1 to regulate the expression of FLO genes, which encode adhesions responsible for yeast biofilm formation, FLO gene transcription levels were measured via qRT-PCR. Relative to wild-type S. cerevisiae, the adhesion genes FLO1, 5, and 9 which also demonstrate increased expression in the transcriptome of yeast cells during biofilm formation, but not FLO11, were down-regulated in the MIG1 mutant strain. Additionally, the MIG1 mutant lost a majority of its flocculation ability, which depended on cell-cell adhesions and its slightly invasive growth ability, dependent on cell-substrate adhesion. Deleting FLO1, 5, and 9 decreased biofilm formation on plastics, suggesting these FLO genes contribute to the biofilm formation process alongside FLO11. Moreover, the ethanol tolerance of yeast decreased in the MIG1 deletion mutant as well as the FLO11 deletion mutant, resulting in reduced biofilm formation during fermentation. It remains possible that in the later period of fermentation, when ethanol has accumulated, an over-expression of the FLO1, 5, and 9 genes regulated by MIG1 would enhanced cell-cell adhesions and thus protect cells in the outer layer of biofilms from ethanol, a function primarily dependent on cell-cell adhesions. This work offers a possible explanation for how biofilm formation is regulated during the immobilized fermentation process, and can enhance environmental tolerance in industrial production.

Cucumber metal tolerance protein 7 (CsMTP7) is involved in the accumulation of Fe in mitochondria under Fe excess.

The plant metal tolerance protein family (MTP) includes 12 members that have been classified into three phylogenetically different subgroups - Zn-cation diffusion facilitator (CDF), Fe/Zn-CDF and Mn-CDF - based on their putative metal specificity. To date, only members belonging to the Zn-CDF or Mn-CDF group have been characterized functionally. The plant Fe/Zn-CDF subgroup includes two proteins, MTP6 and MTP7, but their function and metal specificity have not been confirmed. In this study we showed that cucumber CsMTP7 is a highly specific mitochondrial Fe importer that is able to confer yeast tolerance to Fe excess through increased accumulation of Fe in the mitochondria. We also demonstrated that CsMTP7 contributes to the increased accumulation of Fe in the mitochondria of Arabidopsis thaliana protoplasts. The transcripts and mitochondrial levels of CsMTP7 and ferritin - the iron-storing protein - are significantly increased in cucumber roots in response to Fe excess. This finding suggests that CsMTP7 and ferritin work in concert to accumulate Fe in plant mitochondria. As genes that encode orthologous proteins have been identified in phylogenetically distant organisms, including Archaea, cyanobacteria, humans and plants, but not in yeast, we concluded that the MTP7-mediated mitochondrial Fe accumulation may be conserved in the species, and express mitochondrial ferritin for mitochondrial Fe storage.

Response and tolerance of yeast to changing environmental stress during ethanol fermentation

Abstract In development of yeast cell factories, the adequate stress tolerance of the yeast strains is the universal requirement for industrial fermentation. Primary stress factor is the end product toxicity which has to be accumulated to obtain economically feasible performance in industrial processes. Second stress factor is the concentration and composition of substrate used for ethanol production and the third stress factor is temperature gradients in the fermentor which influence fermentation rate and productivity. This review encloses the development in understanding the response of yeast towards various environmental stresses: heat shock, ethanol stress and osmotic stress. These stresses are fundamental challenges that are experienced by various microbes. Physiological changes such as the synthesis of compatible solutes and heat shock proteins also contribute to cellular survival. Different methods are available to improve the strain’s survival under different stresses; thus, it will not only grow but also produce beneficial products in harsh environments. In combination with rational metabolic engineering to study and analyze processes of adaptive evolution, evolutionary engineering is a powerful strategy for improving industrially important, complex properties of yeast. Evolutionary engineering is an emerging field that draws on the most recent advances from a wide range of scientific and technical disciplines.

Low-pH production of d-lactic acid using newly isolated acid tolerant yeast Pichia kudriavzevii NG7.

Lactic acid is a platform chemical for the sustainable production of various materials. To develop a robust yeast platform for low-pH production of d-lactic acid (LA), an acid-tolerant yeast strain was isolated from grape skins and named Pichia kudriavzevii NG7 by ribosomal RNA sequencing. This strain could grow at pH 2.0 and 50°C. For the commercial application of P. kudriavzevii NG7 as a lactic acid producer, the ethanol fermentation pathway was redirected to lactic acid by replacing the pyruvate decarboxylase 1 gene (PDC1) with the d-lactate dehydrogenase gene (d-LDH) derived from Lactobacillus plantarum. To enhance lactic acid tolerance, this engineered strain was adapted to high lactic acid concentrations, and a new transcriptional regulator, PAR1, responsible for acid tolerance, was identified by whole-genome resequencing. The final engineered strain produced 135 g/L and 154 g/L of d-LA with productivity over 3.66 g/L/hr at pH 3.6 and 4.16 g/L/hr at pH 4.7, respectively.

Antarctic yeasts: analysis of their freeze-thaw tolerance and production of antifreeze proteins, fatty acids and ergosterol

BackgroundMicroorganisms have evolved a number of mechanisms to thrive in cold environments, including the production of antifreeze proteins, high levels of polyunsaturated fatty acids, and ergosterol. In this work, several yeast species isolated from Antarctica were analyzed with respect to their freeze-thaw tolerance and production of the three abovementioned compounds, which may also have economic importance.ResultsThe freeze-thaw tolerance of yeasts was widely variable among species, and a clear correlation with the production of any of the abovementioned compounds was not observed. Antifreeze proteins that were partially purified from Goffeauzyma gastrica maintained their antifreeze activities after several freeze-thaw cycles. A relatively high volumetric production of ergosterol was observed in the yeasts Vishniacozyma victoriae, G. gastrica and Leucosporidium creatinivorum, i.e., 19, 19 and 16 mg l− 1, respectively. In addition, a high percentage of linoleic acid with respect to total fatty acids was observed in V. victoriae (10%), Wickerhamomyces anomalus (12%) and G. gastrica (13%), and a high percentage of alpha linoleic acid was observed in L. creatinivorum (3.3%).ConclusionsGiven these results, the abovementioned yeasts are good candidates to be evaluated for use in the production of antifreeze proteins, fatty acids, and ergosterol at the industrial scale.

The Hog1 positive regulated YCT1 gene expression under cadmium tolerance of budding yeast.

Cadmium (Cd) is a heavy metal that is the cause of irreversible hazards to living organisms. Cadmium ions can induce the phosphorylation of MAPKs pathway molecules such as Hog1 and Slt2, but downstream effectors and potential activation pathways are still unclear. In this study, the RNA-seq data analysis in Cd-stressed yeast was performed to predict and screen the signal transduction pathway and the potential effect molecules regulated by MAPKs. Based on differentially expressed genes and Venn diagrams, 31 genes regulated by Hog1p and two genes induced by Slt2p, which related to carbohydrate metabolism, oxidative damage, DNA replication stress and detoxification, were characterized under Cd exposure to yeast. A cysteine-specific transporter (Yct1) modulated by Hog1 was confirmed via RNA-seq results. Meanwhile, we tested the Cd-sensitivity, intracellular Cd concentrations and β-galactosidase assay, and results indicated that the hypersensitivity of the hog1 mutant to Cd was partly abrogated in YCT1 gene deletion, induction of YCT1 was dependent on Hog1 and its transcription factors, and Yct1p would be epistatic to the Hog1p in Cd-tolerance. The investigation of the transcriptome of MAPKs under Cd stress provided valuable information for future molecular studies of Cd-tolerance.

Mechanism and kinetics of degrading aflatoxin B1 by salt tolerant Candida versatilis CGMCC 3790

Four products were identified by liquid chromatography/time-of-flight mass spectrometry (LC/TOF-MS) for the degradation of aflatoxin B1 (AFB1) is by salt tolerant Candida versatilis CGMCC 3790 (C. versatilis CGMCC 3790), includingⅡ(C14H10O4), Ⅲ (C14H12O3), Ⅳ (C13H12O2), Ⅴ (C11H10O4), which were not toxic. Based on these products, it is speculated that AFB1 degradation has two pathways. The degradation ratio of active cell component (69.40%) and intracellular component (64.99%) was significantly higher than extracellular component (29.61%), suggesting that the AFB1 removal mainly resulted from biodegradation. The optimal degradation conditions of AFB1 (20 ng/mL) were: incubated at pH 5.0, 25 °C for 60 min in liquid medium system. The degradation ratio was ranged from 41.23%∼100% at 10.26∼130.44 ng/g in an actual system. This is the first report revealing that a salt tolerant yeast could effectively degrade AFB1. Therefore, Candida versatilis CGMCC 3790 might be an excellent candidate for bioremediation and detoxification for oriental fermentation condiment process.

The global regulator IrrE from Deinococcus radiodurans enhances the furfural tolerance of Saccharomyces cerevisiae

High tolerance to inhibitors is crucial for microorganisms and difficult to achieve by manipulating single genes because strain tolerance involves multiple genes and several complicated regulation mechanisms. The present study demonstrated that irrE gene from Deinococcus radiodurans R1 promoted the furfural resistance of Saccharomyces cerevisiae through directed evolution. Compared to the wild-type strain, the FR mutant not only showed improved tolerance to single- or multi- inhibitors (furfural, 5-hydroxymethylfurfural, formic and acetic acid), but also obtained better ability of eliminating reactive oxygen species. Consequently, batch fermentation of the FR mutant in synthetic medium (200 g/L glucose and 2.0 g/L furfural) resulted in an 37% increase in ethanol productivity, although ethanol titer (∼88 g/L) was comparable to that of the control. This study provides the first evidence that an exogenous global regulator can improve the stress tolerance of yeast, which is significant for production of industrial products from lignocellulose by S. cerevisiae.

Understanding the tolerance of the industrial yeast Saccharomyces cerevisiae against a major class of toxic aldehyde compounds.

Development of the next-generation biocatalyst is vital for fermentation-based industrial applications and a sustainable bio-based economy. Overcoming the major class of toxic compounds associated with lignocellulose-to-biofuels conversion is one of the significant challenges for new strain development. A significant number of investigations have been made to understand mechanisms of the tolerance for industrial yeast. It is humbling to learn how complicated the cell's response to the toxic chemicals is and how little we have known about yeast tolerance in the universe of the living cell. This study updates our current knowledge on the tolerance of industrial yeast against aldehyde inhibitory compounds at cellular, molecular and the genomic levels. It is comprehensive yet specific based on reproducible evidence and cross confirmed findings from different investigations using varied experimental approaches. This research approaches a rational foundation toward a more comprehensive understanding on the yeast tolerance. Discussions and perspectives are also proposed for continued exploring the puzzle of the yeast tolerance to aid the next-generation biocatalyst development.

Experimental Evolution of Yeast for High-Temperature Tolerance.

Thermotolerance is a polygenic trait that contributes to cell survival and growth under unusually high temperatures. Although some genes associated with high-temperature growth (Htg+) have been identified, how cells accumulate mutations to achieve prolonged thermotolerance is still mysterious. Here, we conducted experimental evolution of a Saccharomyces cerevisiae laboratory strain with stepwise temperature increases for it to grow at 42 °C. Whole genome resequencing of 14 evolved strains and the parental strain revealed a total of 153 mutations in the evolved strains, including single nucleotide variants, small INDELs, and segmental duplication/deletion events. Some mutations persisted from an intermediate temperature to 42 °C, so they might be Htg+ mutations. Functional categorization of mutations revealed enrichment of exonic mutations in the SWI/SNF complex and F-type ATPase, pointing to their involvement in high-temperature tolerance. In addition, multiple mutations were found in a general stress-associated signal transduction network consisting of Hog1 mediated pathway, RAS-cAMP pathway, and Rho1-Pkc1 mediated cell wall integrity pathway, implying that cells can achieve Htg+ partly through modifying existing stress regulatory mechanisms. Using pooled segregant analysis of five Htg+ phenotype-orientated pools, we inferred causative mutations for growth at 42 °C and identified those mutations with stronger impacts on the phenotype. Finally, we experimentally validated a number of the candidate Htg+ mutations. This study increased our understanding of the genetic basis of yeast tolerance to high temperature.

Metabolic and genomic characterisation of stress-tolerant industrial Saccharomyces cerevisiae strains from TALENs-assisted multiplex editing.

TALENs-assisted multiplex editing (TAME) toolbox was previously established and used to successfully enhance ethanol stress tolerance of Saccharomyces cerevisiae laboratory strain. Here, the TAME toolbox was harnessed to improve and elucidate stress tolerances of S. cerevisiae industrial strain. One osmotolerant strain and one thermotolerant strain were selected from the mutant library generated by TAME at corresponding stress conditions, and exhibited 1.2-fold to 1.3-fold increases of fermentation capacities, respectively. Genome resequencing uncovered genomic alterations in the selected stress-tolerant strains, suggesting that cell wall and membrane-related proteins might be major factors behind improved tolerances of yeast to different stresses. Furthermore, amplified mitochondrial DNA might also have an important impact on increased stress tolerance. Unexpectedly, none of predesigned target potential TALENs modification sites showed any genomic variants in sequenced genomes of the selected strains, implicating that the improved stress tolerances might be due to indirect impacts of genome editing via TALENs rather than introducing genomic variants at potential target sites. Our findings not only confirmed TAME could be a useful tool to accelerate the breeding of industrial strain with multiple stress tolerance, but also supported the previous understandings of the complicated mechanisms of multiple stress tolerance in yeast.

Can Zymomonas mobilis Substitute Saccharomyces cerevisiae in Cereal Dough Leavening?

Baker’s yeast intolerance is rising among Western populations, where Saccharomyces cerevisiae is spread in fermented food and food components. Zymomonas mobilis is a bacterium commonly used in tropical areas to produce alcoholic beverages, and it has only rarely been considered for dough leavening probably because it only ferments glucose, fructose and sucrose, which are scarcely present in flour. However, through alcoholic fermentation, similarly to S. cerevisiae, it provides an equimolar mixture of ethanol and CO2 that can rise a dough. Here, we propose Z. mobilis as a new leavening agent, as an alternative to S. cerevisiae, overcoming its technological limit with different strategies: (1) adding glucose to the dough formulation; and (2) exploiting the maltose hydrolytic activity of Lactobacillus sanfranciscensis associated with Z. mobilis. CO2 production, dough volume increase, pH value, microbial counts, sugars consumption and ethanol production were monitored. Results suggest that glucose addition to the dough lets Z. mobilis efficiently leaven a dough, while glucose released by L. sanfranciscensis is not so well fermented by Z. mobilis, probably due to the strong acidification. Nevertheless, the use of Z. mobilis as a leavening agent could contribute to increasing the variety of baked goods alternative to those leavened by S. cerevisiae.

Genomic Sequence of Saccharomyces cerevisiae BAW-6, a Yeast Strain Optimal for Brewing Barley Shochu.

ABSTRACTHere, we report the draft genome sequence of Saccharomyces cerevisiae strain BAW-6, which is used for the production of barley shochu, a traditional Japanese spirit. This genomic information can be used to elucidate the genetic basis underlying the high alcohol production capacity and citric acid tolerance of shochu yeast.

HAA1 and PRS3 overexpression boosts yeast tolerance towards acetic acid improving xylose or glucose consumption: unravelling the underlying mechanisms.

Acetic acid tolerance and xylose consumption are desirable traits for yeast strains used in industrial biotechnological processes. In this work, overexpression of a weak acid stress transcriptional activator encoded by the gene HAA1 and a phosphoribosyl pyrophosphate synthetase encoded by PRS3 in a recombinant industrial Saccharomyces cerevisiae strain containing a xylose metabolic pathway was evaluated in the presence of acetic acid in xylose- or glucose-containing media. HAA1 or PRS3 overexpression resulted in superior yeast growth and higher sugar consumption capacities in the presence of 4g/L acetic acid, and a positive synergistic effect resulted from the simultaneous overexpression of both genes. Overexpressing these genes also improved yeast adaptation to a non-detoxified hardwood hydrolysate with a high acetic acid content. Furthermore, the overexpression of HAA1 and/or PRS3 was found to increase the robustness of yeast cell wall when challenged with acetic acid stress, suggesting the involvement of the modulation of the cell wall integrity pathway. This study clearly shows HAA1 and/or, for the first time, PRS3 overexpression to play an important role in the improvement of industrial yeast tolerance towards acetic acid. The results expand the molecular toolbox and add to the current understanding of the mechanisms involved in higher acetic acid tolerance, paving the way for the further development of more efficient industrial processes.

Yeast and Cancer: Common Mechanism Underlying Activation of Ras by Glycolytic Flux

Ras proteins are small guanosine triphosphatases (GTPases), which function as on-off binary switches for diverse signaling pathways. Nearly 30% of human tumors have mutated forms of Ras. Both in the yeast Saccharomyces cerevisiae and in mammalian cells, Ras is an important regulator of cell proliferation. Mammalian Ras can functionally replace yeast Ras and hyperactive Ras oncogenes cause aberrant cell proliferation in yeast. Also the Ras regulatory proteins, guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs), are homologous between yeast and mammalian cells. We used the tps1Δ mutant of the yeast S. cerevisiae as a tool to study Ras regulation. In the presence of glucose, this strain cannot grow due to a major metabolic deregulation causing hyperaccumulation of fructose-1,6-bisphosphate (Fru-1,6-bisP) and also hyperactivation of Ras leading to apoptosis. This led us to the finding that upon addition of glucose to wild type yeast cells, Fru-1,6-bisP promotes - in physiological concentrations - the activation of Ras through the GEF factor Cdc25. This activation may also be relevant in tumor biology because of the high glycolytic flux and higher concentrations of glycolytic intermediates present in cancer cells (Warburg effect). In vitro studies showed that Fru-1,6-bisP helps to increase the dissociation rate of the human K-Ras/SOS1 complex. Furthermore, we were able to show that glucose addition to glucose-deprived HEK293T and Hela Kyoto cells triggers rapid activation of Ras, as well as activation of its downstream targets MEK and ERK. Our results suggest that there may be a causal link between the high glycolytic activity and rapid cell proliferation of yeast and cancer cells in glucose medium through Fru-1,6-bisP activation of Ras. Support or Funding Information This work was supported by a PhD fellowship from the Agency for Innovation by Science and Technology (IWT-Flanders) to K.P., F.V.L., D.H., and W.V., by a post- doctoral F+-fellowship from the Research Fund of the KU Leuven to H.Q., and by grants from the Fund for Scientific Research—Flanders, Interuniversity Attraction Poles Net- work P6/14 and P7/40, and the Research Fund of the KU Leuven (Concerted Research Actions) to J.M.T., and a grant from the Belgian Foundation Against Cancer (FAF-F/ 2016/822) to V.J. Schematic overview of initial glucose metabolism in yeast and its connection to activation of the Ras-cAMP-PKA pathway. This pathway exerts major control over cell proliferation, fermentation rate, stress tolerance and developmental pathways in yeast. We show that Fru1,6bisP functions as activator of Ras by interacting with Cdc25 and that this connection between glycolysis and Ras is conserved between yeast and mammalian cells. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The Rim Pathway Mediates Antifungal Tolerance in Candida albicans through Newly Identified Rim101 Transcriptional Targets, Including Hsp90 and Ipt1.

Invasive candidiasis (IC) is a major cause of morbidity and mortality despite antifungal treatment. Azoles and echinocandins are used as first-line therapies for IC. However, their efficacy is limited by yeast tolerance and the emergence of acquired resistance. Tolerance is a reversible stage created due to the yeast's capacity to counter antifungal drug exposure, leading to persistent growth. For Candida albicans, multiple stress signaling pathways have been shown to contribute to this adaptation. Among them, the pH-responsive Rim pathway, through its transcription factor Rim101p, was shown to mediate azole and echinocandin tolerance. The Rim pathway is fungus specific, is conserved among the members of the fungal kingdom, and plays a key role in pathogenesis and virulence. The present study aimed at confirming the role of Rim101p and investigating the implication of the other Rim proteins in antifungal tolerance in C. albicans, as well as the mechanisms underlying it. Time-kill curve experiments and colony formation tests showed that genetic inhibition of all the Rim factors enhances echinocandin and azole antifungal activity. Through RNA sequencing analysis of a rim101-/- mutant, a strain constitutively overexpressing RIM101, and control strains, we discovered novel Rim-dependent genes involved in tolerance, including HSP90, encoding a major molecular chaperone, and IPT1, involved in sphingolipid biosynthesis. Rim mutants were also hypersensitive to pharmacological inhibition of Hsp90. Taken together, these data suggest that Rim101 acts upstream of Hsp90 and that targeting the Rim pathway in combination with existing antifungal drugs may represent a promising antifungal strategy to indirectly but specifically target Hsp90 in yeasts.

Industrial Applications of Selected JFS Articles

Industrial Applications of Selected <i>JFS</i> Articles

Selection of Yeast Strains for Tequila Fermentation Based on Growth Dynamics in Combined Fructose and Ethanol Media.

A method to select yeast strains for fermentation taking into account the natural differences of yeast isolates. This methodology is based on the cell exposition to combinations of sugar and ethanol, which are the most important stress factors in fermentation. This strategy will help to identify the most tolerant strain that could improve ethanol yield and reduce fermentation time.

Chemical genomic guided engineering of gamma-valerolactone tolerant yeast

BackgroundGamma valerolactone (GVL) treatment of lignocellulosic bomass is a promising technology for degradation of biomass for biofuel production; however, GVL is toxic to fermentative microbes. Using a combination of chemical genomics with the yeast (Saccharomyces cerevisiae) deletion collection to identify sensitive and resistant mutants, and chemical proteomics to monitor protein abundance in the presence of GVL, we sought to understand the mechanism toxicity and resistance to GVL with the goal of engineering a GVL-tolerant, xylose-fermenting yeast.ResultsChemical genomic profiling of GVL predicted that this chemical affects membranes and membrane-bound processes. We show that GVL causes rapid, dose-dependent cell permeability, and is synergistic with ethanol. Chemical genomic profiling of GVL revealed that deletion of the functionally related enzymes Pad1p and Fdc1p, which act together to decarboxylate cinnamic acid and its derivatives to vinyl forms, increases yeast tolerance to GVL. Further, overexpression of Pad1p sensitizes cells to GVL toxicity. To improve GVL tolerance, we deleted PAD1 and FDC1 in a xylose-fermenting yeast strain. The modified strain exhibited increased anaerobic growth, sugar utilization, and ethanol production in synthetic hydrolysate with 1.5% GVL, and under other conditions. Chemical proteomic profiling of the engineered strain revealed that enzymes involved in ergosterol biosynthesis were more abundant in the presence of GVL compared to the background strain. The engineered GVL strain contained greater amounts of ergosterol than the background strain.ConclusionsWe found that GVL exerts toxicity to yeast by compromising cellular membranes, and that this toxicity is synergistic with ethanol. Deletion of PAD1 and FDC1 conferred GVL resistance to a xylose-fermenting yeast strain by increasing ergosterol accumulation in aerobically grown cells. The GVL-tolerant strain fermented sugars in the presence of GVL levels that were inhibitory to the unmodified strain. This strain represents a xylose fermenting yeast specifically tailored to GVL produced hydrolysates.

Abiotic stress response in yeast of a type 1 alternative oxidase (LpAOX1) from Lilium pumilum

We cloned an LpAOX1 gene from Lilium puminlum , explored this gene expression in yeast under salts (NaCl, Na 2 CO 3 , NaHCO 3 ), pH value or oxidation (H 2 O 2 ) stress, the transgenic yeast growth better than untransgenic yeast in presence of stresses. The result reveal that LpAOX1 gene has effect on the yeast tolerance to the stresses and contribute to the elucidation of the LpAOX1 molecular mechanisms involved in the stress response pathway. LpAOX1 expression in the yeast control harmful ROS contents generation, alleviate the ROS induced oxidative stress and the resistance of transgenic yeast to salt stress was enhanced.