scholarly journals Wine Yeast Peroxiredoxin TSA1 Plays a Role in Growth, Stress Response and Trehalose Metabolism in Biomass Propagation

2020 ◽  
Vol 8 (10) ◽  
pp. 1537
Author(s):  
Víctor Garrigós ◽  
Cecilia Picazo ◽  
Emilia Matallana ◽  
Agustín Aranda
Keyword(s):  
Gene Deletion ◽  
Carbon Sources ◽  
Thioredoxin Reductase ◽  
Reducing Power ◽  
Grape Juice ◽  
Normal Growth ◽  
Wine Yeast ◽  
Growth Stress ◽  
Growth Stages ◽  
Glycogen Accumulation

Peroxiredoxins are a family of peroxide-degrading enzymes for challenging oxidative stress. They receive their reducing power from redox-controlling proteins called thioredoxins, and these, in turn, from thioredoxin reductase. The main cytosolic peroxiredoxin is Tsa1, a moonlighting protein that also acts as protein chaperone a redox switch controlling some metabolic events. Gene deletion of peroxiredoxins in wine yeasts indicate that TSA1, thioredoxins and thioredoxin reductase TRR1 are required for normal growth in medium with glucose and sucrose as carbon sources. TSA1 gene deletion also diminishes growth in molasses, both in flasks and bioreactors. The TSA1 mutation brings about an expected change in redox parameters but, interestingly, it also triggers a variety of metabolic changes. It influences trehalose accumulation, lowering it in first molasses growth stages, but increasing it at the end of batch growth, when respiratory metabolism is set up. Glycogen accumulation at the entry of the stationary phase also increases in the tsa1Δ mutant. The mutation reduces fermentative capacity in grape juice, but the vinification profile does not significantly change. However, acetic acid and acetaldehyde production decrease when TSA1 is absent. Hence, TSA1 plays a role in the regulation of metabolic reactions leading to the production of such relevant enological molecules.

scholarly journals Comparative genomics in the wine bacterium Oenococcus oeni

2011 ◽  
Vol 32 (4) ◽  
pp. 174
Author(s):  
Anthony R Borneman ◽  
Eveline J Bartowsky
Keyword(s):  
Carbon Sources ◽  
Grape Juice ◽  
Wine Yeast ◽  
Oenococcus Oeni ◽  
Primary Role ◽  
Yeast Saccharomyces Cerevisiae ◽  
Spoilage Bacteria ◽  
Microbial Stability ◽  
Mouth Feel ◽  
By Products

The production of wine from grape juice relies on the combined actions of both yeast and bacteria which shape the aroma and flavour of wine through the production of secondary metabolites and the biochemical transformation of many grape-derived constituents. Whereas the principal wine yeast, Saccharomyces cerevisiae, is primarily involved in the alcoholic fermentation in which glucose and fructose are converted into alcohol, the wine bacterium, Oenococcus oeni, is primarily involved in a secondary fermentation reaction where malic acid is decarboxlyated into lactic acid. This conversion, known as malolactic fermentation (MLF), results in an increase in wine pH and reduction in the sourness of the wine, while also providing microbial stability through the reduction of potential carbon sources for wine spoilage bacteria such as Lactobacilli and Pediococci1. In addition to its primary role in performing MLF, the metabolic by-products produced during the growth of O. oeni in wine have been shown to positively contribute to the flavour and mouth feel of wines which have undergone MLF.

scholarly journals Towards Initial Indications for a Thiol-Based Redox Control of Arabidopsis 5-Aminolevulinic Acid Dehydratase

Antioxidants ◽  
2018 ◽  
Vol 7 (11) ◽  
pp. 152 ◽  
Author(s):  
Daniel Wittmann ◽  
Sigri Kløve ◽  
Peng Wang ◽  
Bernhard Grimm
Keyword(s):  
Aminolevulinic Acid ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Thioredoxin Reductase ◽  
Reducing Power ◽  
Reduced Form ◽  
Redox Control ◽  
In Planta ◽  
Acid Dehydratase ◽  
Reducing Conditions ◽  
Alad Activity

Thiol-based redox control is one of the important posttranslational mechanisms of the tetrapyrrole biosynthesis pathway. Many enzymes of the pathway have been shown to interact with thioredoxin (TRX) and Nicotinamide adenine dinucleotide phosphate (NADPH)-dependent thioredoxin reductase C (NTRC). We examined the redox-dependency of 5-aminolevulinic acid dehydratase (ALAD), which catalyzed the conjugation of two 5-aminolevulinic acid (ALA) molecules to porphobilinogen. ALAD interacted with TRX f, TRX m and NTRC in chloroplasts. Consequently, less ALAD protein accumulated in the trx f1, ntrc and trx f1/ntrc mutants compared to wild-type control resulting in decreased ALAD activity. In a polyacrylamide gel under non-reducing conditions, ALAD monomers turned out to be present in reduced and two oxidized forms. The reduced and oxidized forms of ALAD differed in their catalytic activity. The addition of TRX stimulated ALAD activity. From our results it was concluded that (i) deficiency of the reducing power mainly affected the in planta stability of ALAD; and (ii) the reduced form of ALAD displayed increased enzymatic activity.

scholarly journals Interplay between cytosolic disulfide reductase systems and the Nrf2/Keap1 pathway

2015 ◽  
Vol 43 (4) ◽  
pp. 632-638 ◽  
Author(s):  
Edward E. Schmidt
Keyword(s):  
Oxidative Stress ◽  
Thioredoxin Reductase ◽  
Reducing Power ◽  
Nuclear Factor ◽  
Independent Source ◽  
Nrf2 Pathway ◽  
Thioredoxin Reductase 1 ◽  
Disulfide Reductase ◽  
The Status ◽  
Thioredoxin 1

NADPH transfers reducing power from bioenergetic pathways to thioredoxin reductase-1 (TrxR1) and glutathione reductase (GR) to support essential reductive systems. Surprisingly, it was recently shown that mouse livers lacking both TrxR1 and GR (‘TR/GR-null’) can sustain redox (reduction-oxidation) homoeostasis using a previously unrecognized NADPH-independent source of reducing power fuelled by dietary methionine. The NADPH-dependent systems are robustly redundant in liver, such that disruption of either TrxR1 or GR alone does not cause oxidative stress. However, disruption of TrxR1 induces transcription factor Nrf2 (nuclear factor erythroid-derived 2-like-2) whereas disruption of GR does not. This suggests the Nrf2 pathway responds directly to the status of the thioredoxin-1 (Trx1) system. The proximal regulator of Nrf2 is Keap1 (Kelch-like ECH-associated protein-1), a cysteine (Cys)-rich protein that normally interacts transiently with Nrf2, targeting it for degradation. During oxidative stress, this interaction is stabilized, preventing degradation of newly synthesized Nrf2, thereby allowing Nrf2 accumulation. Within the Trx1 system, TrxR1 and peroxiredoxins (Prxs) contain some of the most reactive nucleophilic residues in the cell, making them likely targets for oxidants or electrophiles. We propose that Keap1 activity and therefore Nrf2 is regulated by interactions of Trx1 system enzymes with oxidants. In TR/GR-null livers, Nrf2 activity is further induced, revealing that TrxR-independent systems also repress Nrf2 and these might be induced by more extreme challenges.

scholarly journals YvcK of Bacillus subtilis is required for a normal cell shape and for growth on Krebs cycle intermediates and substrates of the pentose phosphate pathway

Microbiology ◽  
2005 ◽  
Vol 151 (11) ◽  
pp. 3777-3791 ◽  
Author(s):  
Boris Görke ◽  
Elodie Foulquier ◽  
Anne Galinier
Keyword(s):  
Bacillus Subtilis ◽  
Pentose Phosphate Pathway ◽  
Carbon Metabolism ◽  
Carbon Sources ◽  
Krebs Cycle ◽  
Normal Growth ◽  
Central Carbon Metabolism ◽  
Pentose Phosphate ◽  
Homologous Protein ◽  
Krebs Cycle Intermediates

The HPr-like protein Crh has so far been detected only in the bacillus group of bacteria. In Bacillus subtilis, its gene is part of an operon composed of six ORFs, three of which exhibit strong similarity to genes of unknown function present in many bacteria. The promoter of the operon was determined and found to be constitutively active. A deletion analysis revealed that gene yvcK, encoded by this operon, is essential for growth on Krebs cycle intermediates and on carbon sources metabolized via the pentose phosphate pathway. In addition, cells lacking YvcK acquired media-dependent filamentous or L-shape-like aberrant morphologies. The presence of high magnesium concentrations restored normal growth and cell morphology. Furthermore, suppressor mutants cured from these growth defects appeared spontaneously with a high frequency. Such suppressing mutations were identified in a transposon mutagenesis screen and found to reside in seven different loci. Two of them mapped in genes of central carbon metabolism, including zwf, which encodes glucose-6-phosphate dehydrogenase and cggR, the product of which regulates the synthesis of glyceraldehyde-3-phosphate dehydrogenase. All these results suggest that YvcK has an important role in carbon metabolism, probably in gluconeogenesis required for the synthesis of cell wall precursor molecules. Interestingly, the Escherichia coli homologous protein, YbhK, can substitute for YvcK in B. subtilis, suggesting that the two proteins have been functionally conserved in these different bacteria.

scholarly journals Long-Term Adaption to High Osmotic Stress as a Tool for Improving Enological Characteristics in Industrial Wine Yeast

Genes ◽  
2020 ◽  
Vol 11 (5) ◽  
pp. 576 ◽  
Author(s):  
Gabriela Betlej ◽  
Ewelina Bator ◽  
Bernadetta Oklejewicz ◽  
Leszek Potocki ◽  
Anna Górka ◽  
...  
Keyword(s):  
Osmotic Stress ◽  
Grape Juice ◽  
Wine Yeast ◽  
Sensory Profile ◽  
Wine Yeasts ◽  
Adaptive Laboratory Evolution ◽  
Yeast Viability ◽  
Glycerol Synthesis ◽  
The Many

Industrial wine yeasts owe their adaptability in constantly changing environments to a long evolutionary history that combines naturally occurring evolutionary events with human-enforced domestication. Among the many stressors associated with winemaking processes that have potentially detrimental impacts on yeast viability, growth, and fermentation performance are hyperosmolarity, high glucose concentrations at the beginning of fermentation, followed by the depletion of nutrients at the end of this process. Therefore, in this study, we subjected three widely used industrial wine yeasts to adaptive laboratory evolution under potassium chloride (KCl)-induced osmotic stress. At the end of the evolutionary experiment, we evaluated the tolerance to high osmotic stress of the evolved strains. All of the analyzed strains improved their fitness under high osmotic stress without worsening their economic characteristics, such as growth rate and viability. The evolved derivatives of two strains also gained the ability to accumulate glycogen, a readily mobilized storage form of glucose conferring enhanced viability and vitality of cells during prolonged nutrient deprivation. Moreover, laboratory-scale fermentation in grape juice showed that some of the KCl-evolved strains significantly enhanced glycerol synthesis and production of resveratrol-enriched wines, which in turn greatly improved the wine sensory profile. Altogether, these findings showed that long-term adaptations to osmotic stress can be an attractive approach to develop industrial yeasts.

scholarly journals Alterations in Yeast Species Composition of Uninoculated Wine Ferments by the Addition of Sulphur Dioxide

Fermentation ◽  
2020 ◽  
Vol 6 (2) ◽  
pp. 62 ◽  
Author(s):  
Kathleen Cuijvers ◽  
Steven Van Den Heuvel ◽  
Cristian Varela ◽  
Mark Rullo ◽  
Mark Solomon ◽  
...  
Keyword(s):  
Community Structure ◽  
Sulphur Dioxide ◽  
Yeast Species ◽  
Grape Juice ◽  
Wine Yeast ◽  
Aroma Compound ◽  
Microbial Composition ◽  
Yeast Community ◽  
Selection For ◽  
Saccharomyces Yeasts

Uninoculated wine fermentations are conducted by a consortium of wine yeast and bacteria that establish themselves either from the grape surface or from the winery environment. Of the additives that are commonly used by winemakers, sulphur dioxide (SO2) represents the main antimicrobial preservative and its use can have drastic effects on the microbial composition of the fermentation. To investigate the effect of SO2 on the resident yeast community of uninoculated ferments, Chardonnay grape juice from 2018 and 2019 was treated with a variety of SO2 concentrations ranging up to 100 mg/L and was then allowed to undergo fermentation, with the yeast community structure being assessed via high-throughput meta-barcoding (phylotyping). While the addition of SO2 was shown to select against the presence of many species of non-Saccharomyces yeasts, there was a clear and increasing selection for the species Hanseniaspora osmophila as concentrations of SO2 rose above 40 mg/L in fermentations from both vintages. Chemical analysis of the wines resulting from these treatments showed significant increases in acetate esters, and specifically the desirable aroma compound 2-phenylethyl acetate, that accompanied the increase in abundance of H. osmophila. The ability to modulate the yeast community structure of an uninoculated ferment and the resulting chemical composition of the final wine, as demonstrated in this study, represents an important tool for winemakers to begin to be able to influence the organoleptic profile of uninoculated wines.

Genome-scale metabolic modeling to provide insight into the production of storage compounds during feast–famine cycles of activated sludge

2013 ◽  
Vol 67 (3) ◽  
pp. 469-476 ◽  
Author(s):  
Mohammad Tajparast ◽  
Dominic Frigon
Keyword(s):  
Activated Sludge ◽  
Carbon Sources ◽  
Value Added ◽  
Growth Cycle ◽  
Glycogen Accumulation ◽  
Storage Compounds ◽  
Metabolic Models ◽  
K 12 ◽  
Genome Scale ◽  
By Products

Studying storage metabolism during feast–famine cycles of activated sludge treatment systems provides profound insight in terms of both operational issues (e.g., foaming and bulking) and process optimization for the production of value added by-products (e.g., bioplastics). We examined the storage metabolism (including poly-β-hydroxybutyrate [PHB], glycogen, and triacylglycerols [TAGs]) during feast–famine cycles using two genome-scale metabolic models: Rhodococcus jostii RHA1 (iMT1174) and Escherichia coli K-12 (iAF1260) for growth on glucose, acetate, and succinate. The goal was to develop the proper objective function (OF) for the prediction of the main storage compound produced in activated sludge for given feast–famine cycle conditions. For the flux balance analysis, combinations of three OFs were tested. For all of them, the main OF was to maximize growth rates. Two additional sub-OFs were used: (1) minimization of biochemical fluxes, and (2) minimization of metabolic adjustments (MoMA) between the feast and famine periods. All (sub-)OFs predicted identical substrate–storage associations for the feast–famine growth of the above-mentioned metabolic models on a given substrate when glucose and acetate were set as sole carbon sources (i.e., glucose–glycogen and acetate–PHB), in agreement with experimental observations. However, in the case of succinate as substrate, the predictions depended on the network structure of the metabolic models such that the E. coli model predicted glycogen accumulation and the R. jostii model predicted PHB accumulation. While the accumulation of both PHB and glycogen was observed experimentally, PHB showed higher dynamics during an activated sludge feast–famine growth cycle with succinate as substrate. These results suggest that new modeling insights between metabolic predictions and population ecology will be necessary to properly predict metabolisms likely to emerge within the niches of activated sludge communities. Nonetheless, we believe that the development of this approach will help guide the optimization of the production of storage compounds as valuable by-products of wastewater treatment.

scholarly journals Exopolysaccharide Production Is Required for Biofilm Formation and Plant Colonization by the Nitrogen-Fixing Endophyte Gluconacetobacter diazotrophicus

2011 ◽  
Vol 24 (12) ◽  
pp. 1448-1458 ◽  
Author(s):  
Carlos H. S. G. Meneses ◽  
Luc F. M. Rouws ◽  
Jean L. Simões-Araújo ◽  
Marcia S. Vidal ◽  
José I. Baldani
Keyword(s):  
Biofilm Formation ◽  
Bacterial Species ◽  
Carbon Sources ◽  
Normal Growth ◽  
Root Surface ◽  
Plant Colonization ◽  
Gluconacetobacter Diazotrophicus ◽  
Nitrogen Fixing ◽  
Wild Type ◽  
Endophytic Colonization

The genome of the endophytic diazotrophic bacterial species Gluconacetobacter diazotrophicus PAL5 (PAL5) revealed the presence of a gum gene cluster. In this study, the gumD gene homologue, which is predicted to be responsible for the first step in exopolysaccharide (EPS) production, was insertionally inactivated and the resultant mutant (MGD) was functionally studied. The mutant MGD presented normal growth and nitrogen (N2) fixation levels but did not produce EPS when grown on different carbon sources. MGD presented altered colony morphology on soft agar plates (0.3% agar) and was defective in biofilm formation on glass wool. Most interestingly, MGD was defective in rice root surface attachment and in root surface and endophytic colonization. Genetic complementation reverted all mutant phenotypes. Also, the addition of EPS purified from culture supernatants of the wild-type strain PAL5 to the mutant MGD was effective in partially restoring wild-type biofilm formation and plant colonization. These data provide strong evidence that the PAL5 gumD gene is involved in EPS biosynthesis and that EPS biosynthesis is required for biofilm formation and plant colonization. To our knowledge, this is the first report of a role of EPS in the endophytic colonization of graminaceous plants by a nitrogen-fixing bacterium.

Distinct electron transfer from ferredoxin–thioredoxin reductase to multiple thioredoxin isoforms in chloroplasts

2017 ◽  
Vol 474 (8) ◽  
pp. 1347-1360 ◽  
Author(s):  
Keisuke Yoshida ◽  
Toru Hisabori
Keyword(s):  
Electron Transfer ◽  
Redox Regulation ◽  
Thioredoxin Reductase ◽  
Reducing Power ◽  
Redox Potentials ◽  
Regulation Network ◽  
Electron Transfers ◽  
Kinetics Of

Thiol-based redox regulation is considered to support light-responsive control of various chloroplast functions. The redox cascade via ferredoxin–thioredoxin reductase (FTR)/thioredoxin (Trx) has been recognized as a key to transmitting reducing power; however, Arabidopsis thaliana genome sequencing has revealed that as many as five Trx subtypes encoded by a total of 10 nuclear genes are targeted to chloroplasts. Because each Trx isoform seems to have a distinct target selectivity, the electron distribution from FTR to multiple Trxs is thought to be the critical branch point for determining the consequence of chloroplast redox regulation. In the present study, we aimed to comprehensively characterize the kinetics of electron transfer from FTR to 10 Trx isoforms. We prepared the recombinant FTR protein from Arabidopsis in the heterodimeric form containing the Fe–S cluster. By reconstituting the FTR/Trx system in vitro, we showed that FTR prepared here was enzymatically active and suitable for uncovering biochemical features of chloroplast redox regulation. A series of redox state determinations using the thiol-modifying reagent, 4-acetamido-4′-maleimidylstilbene-2,2′-disulfonate, indicated that all chloroplast Trx isoforms are commonly reduced by FTR; however, significantly different efficiencies were evident. These differences were apparently correlated with the distinct midpoint redox potentials among Trxs. Even when the experiments were performed under conditions of hypothetical in vivo stoichiometry of FTR and Trxs, a similar trend in distinguishable electron transfers was observed. These data highlight an aspect of highly organized circuits in the chloroplast redox regulation network.

scholarly journals The SSU1 checkup, a rapid tool for detecting chromosomal rearrangements of the Saccharomyces cerevisiae chromosome XVI. An ecological and technological study on wine yeast

2020 ◽  
Author(s):  
Philippe Marullo ◽  
Olivier Claisse ◽  
Maria Laura Raymond Eder ◽  
Marine Börlin ◽  
Nadine Feghali ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Chromosomal Rearrangements ◽  
Decisive Role ◽  
Grape Juice ◽  
Wine Yeast ◽  
Single Experiment ◽  
Pcr Method ◽  
Rapid Characterization ◽  
Technological Study ◽  
And Inversion

1)AbstractChromosomal rearrangements (CR) such as translocations, duplications and inversions play a decisive role in the adaptation of microorganisms to specific environments. In enological Saccharomyces cerevisiae strains, CR involving the promoter region of the gene SSU1 lead to a higher sulfite tolerance by enhancing the SO2 efflux. To date, three different SSU1 associated CR events have been described, including translocations XV-t-XVI and VIII-t-XVI and inversion inv-XVI. In the present study, we developed a multiplex PCR method (SSU1 check-up) that allows a rapid characterization of these three chromosomal configurations in a single experiment. Nearly 600 S. cerevisiae strains collected from fermented grape juice were genotyped by microsatellite markers. We demonstrated that alleles of the SSU1 promoter are differently distributed according to the wine environment (cellar versus vineyard) and the nature of the grape juice. Moreover, rearranged SSU1 promoters are significantly enriched among commercial starters. In addition, nearly isogenic strains collected in similar environments show different CR suggesting that translocation events occur with a non-negligible frequency in clonal populations likely due to mitotic recombination events. Finally, the link between the nature of SSU1 promoter and the tolerance to sulfite was statistically validated in natural grape juice containing various SO2 concentrations. The SSU1 check-up is therefore a convenient new tool for addressing population genetics questions and for selecting yeast strains by using molecular markers.

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