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Jean Guzzo - One of the best experts on this subject based on the ideXlab platform.
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technological properties of Oenococcus oeni strains isolated from typical southern italian wines
Letters in Applied Microbiology, 2009Co-Authors: Vittorio Capozzi, Pasquale Russo, Luciano Beneduce, Stephanie Weidmann, Francesco Grieco, Jean Guzzo, Giuseppe SpanoAbstract:Aims: To isolate indigenous Oenococcus oeni strains suitable as starters for malolactic fermentation (MLF), using a reliable polyphasic approach. Methods and Results: Oenococcus oeni strains were isolated from Nero di Troia wines undergoing spontaneous MLF. Samples were taken at the end of alcoholic fermentation and during MLF. Wine samples were diluted in a sterile physiological solution and plated on MRS and on modified FT80. Identification of O. oeni strains was performed by a polymerase chain reaction (PCR) experiment using strain-specific primers. Strains were further grouped using a multiplex RAPD-PCR analysis. Then, six strains were inoculated in two wine-like media with two different ethanol concentrations (11 and 13% vol/vol) with a view to evaluate their capacity to grow and to perform MLF. In addition, a quantitative PCR (qRT-PCR) approach was adapted to monitor the physiological state of the strains selected. Conclusion: A positive correlation between the malolactic activity performance and the ability to develop and tolerate stress conditions was observed for two selected O. oeni strains. Significance and Impact of the Study: The results reported are useful for the selection of indigenous MLF starter cultures with desired oenological traits from typical regional wines. It should be the base for the improvement in organoleptic quality of typical red wine.
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Oenococcus oeni preference for peptides qualitative and quantitative analysis of nitrogen assimilation
Archives of Microbiology, 2006Co-Authors: Fabienne Remize, Jean Guzzo, Herve Alexandre, Aurelie Gaudin, Yu Kong, Sibylle Krieger, Michele GuillouxbenatierAbstract:Optimization of malolactic fermentation in wine depends mainly on better understanding of nitrogen nutritional requirements of Oenococcus oeni. Four widely used starter strains and the reference ATCC BAA-1163 strain were grown in media containing different N sources: free amino acids, oligopeptides (0.5-10 kDa) or polypeptides (> 10 kDa). Amino acid auxotrophies were determined by the single omission technique. The tested strains were indifferent to only two to four amino acids and two of the starter strains appeared to be particularly demanding. Nitrogen consumption was investigated and a significant level of nitrogen was consumed by O. oeni only in the free amino acid medium. In media containing complex nitrogen sources, a global balance above 5 mg N l(-1) was enough to ensure biomass formation of all tested strains. Moreover, for all strains, bacterial growth yield was higher in the presence of nitrogen from peptides than that from free amino acids. However, no direct relationship between the bacterial growth level and the amount of nitrogen metabolized could be established. These findings were discussed in relation to the physiology of wine malolactic bacteria.
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Real-time PCR for characterizing the stress response of Oenococcus oeni in a wine-like medium
Research in Microbiology, 2006Co-Authors: Charlotte Beltramo, Nicolas Desroche, Raphaëlle Tourdot-maréchal, Cosette Grandvalet, Jean GuzzoAbstract:The tolerance of the lactic acid bacterium Oenococcus oeni to hostile wine conditions is essential for the success of malolactic fermentation (MLF). In this study, reverse transcription quantitative PCR (RT-qPCR) was used to quantify the transcript level of 13 genes that could play a role in adaptation of O. oeni in wine. To optimize survival and growth in wine, cells were adapted during growth at low pH (3.5) prior to inoculation into wine. The level of gene expression was analyzed after growth at pH 3.5 in a rich medium and during MLF in a wine-like medium. RT-qPCR analyses exhibited different expression ratios of stress genes. The data obtained showed that determination of mRNA levels could constitute a new approach to studying the stress response of O. oeni after adaptation at low pH and during growth in a wine-like medium.
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CtsR Is the Master Regulator of Stress Response Gene Expression in Oenococcus oeni
Journal of Bacteriology, 2005Co-Authors: Cosette Grandvalet, Charlotte Beltramo, Françoise Coucheney, Jean GuzzoAbstract:Although many stress response genes have been characterized in Oenococcus oeni, little is known about the regulation of stress response in this malolactic bacterium. The expression of eubacterial stress genes is controlled both positively and negatively at the transcriptional level. Overall, negative regulation of heat shock genes appears to be more widespread among gram-positive bacteria. We recently identified an ortholog of the ctsR gene in O. oeni. In Bacillus subtilis, CtsR negatively regulates expression of the clp genes, which belong to the class III family of heat shock genes. The ctsR gene of O. oeni is cotranscribed with the downstream clpC gene. Sequence analysis of the O. oeni IOB 8413 (ATCC BAA-1163) genome revealed the presence of potential CtsR operator sites upstream from most of the major molecular chaperone genes, including the clp genes and the groES and dnaK operons. Using B. subtilis as a heterologous host, CtsR-dependent regulation of O. oeni molecular chaperone genes was demonstrated with transcriptional fusions. No alternative sigma factors appear to be encoded by the O. oeni IOB 8413 (ATCC BAA-1163) genome. Moreover, apart from CtsR, no known genes encoding regulators of stress response, such as HrcA, could be identified in this genome. Unlike the multiple regulatory mechanisms of stress response described in many closely related gram-positive bacteria, this is the first example where dnaK and groESL are controlled by CtsR but not by HrcA.
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determination of an internal control to apply reverse transcription quantitative pcr to study stress response in the lactic acid bacterium Oenococcus oeni
Journal of Microbiological Methods, 2005Co-Authors: Nicolas Desroche, Charlotte Beltramo, Jean GuzzoAbstract:The expression gene pattern reflects, in part, mechanisms involved in adaptation to environmental conditions. Thus, we established and validated a method that enables relative transcript quantification in different conditions in the lactic acid bacteria Oenococcus oeni, notably in a technological medium. First, we determined an internal control in our conditions by reverse transcription quantitative polymerase chain reaction (RT-qPCR) using the SYBRR Green I technology. Among the seven presumed housekeeping tested genes, the ldhD gene was retained for further experiments. Then, the PCR reproducibility was verified in our conditions and the comparative critical threshold (2 DDC T
Tjakko Abee - One of the best experts on this subject based on the ideXlab platform.
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effect of adaptation to ethanol on cytoplasmic and membrane protein profiles of Oenococcus oeni
Applied and Environmental Microbiology, 2004Co-Authors: Graca M Da Silveira, F M Rombouts, Maja Baumgartner, Tjakko AbeeAbstract:The practical application of commercial malolactic starter cultures of Oenococcus oeni surviving direct inoculation in wine requires insight into mechanisms of ethanol toxicity and of acquired ethanol tolerance in this organism. Therefore, the site-specific location of proteins involved in ethanol adaptation, including cytoplasmic, membrane-associated, and integral membrane proteins, was investigated. Ethanol triggers alterations in protein patterns of O. oeni cells stressed with 12% ethanol for 1 h and those of cells grown in the presence of 8% ethanol. Levels of inosine-5′-monophosphate dehydrogenase and phosphogluconate dehydrogenase, which generate reduced nicotinamide nucleotides, were decreased during growth in the presence of ethanol, while glutathione reductase, which consumes NADPH, was induced, suggesting that maintenance of the redox balance plays an important role in ethanol adaptation. Phosphoenolpyruvate:mannose phosphotransferase system (PTS) components of mannose PTS, including the phosphocarrier protein HPr and EIIMan, were lacking in ethanol-adapted cells, providing strong evidence that mannose PTS is absent in ethanol-adapted cells, and this represents a metabolic advantage to O. oeni cells during malolactic fermentation. In cells grown in the presence of ethanol, a large increase in the number of membrane-associated proteins was observed. Interestingly, two of these proteins, dTDT-glucose-4,6-dehydratase and d-alanine:d-alanine ligase, are known to be involved in cell wall biosynthesis. Using a proteomic approach, we provide evidence for an active ethanol adaptation response of O. oeni at the cytoplasmic and membrane protein levels.
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membrane fluidity adjustments in ethanol stressed Oenococcus oeni cells
Applied and Environmental Microbiology, 2003Co-Authors: Graca M Da Silveira, Elena A Golovina, Folkert A Hoekstra, F M Rombouts, Tjakko AbeeAbstract:The effect of ethanol on the cytoplasmic membrane of Oenococcus oeni cells and the role of membrane changes in the acquired tolerance to ethanol were investigated. Membrane tolerance to ethanol was defined as the resistance to ethanol-induced leakage of preloaded carboxyfluorescein (cF) from cells. To probe the fluidity of the cytoplasmic membrane, intact cells were labeled with doxyl-stearic acids and analyzed by electron spin resonance spectroscopy. Although the effect of ethanol was noticeable across the width of the membrane, we focused on fluidity changes at the lipid-water interface. Fluidity increased with increasing concentrations of ethanol. Cells responded to growth in the presence of 8% (vol/vol) ethanol by decreasing fluidity. Upon exposure to a range of ethanol concentrations, these adapted cells had reduced fluidity and cF leakage compared with cells grown in the absence of ethanol. Analysis of the membrane composition revealed an increase in the degree of fatty acid unsaturation and a decrease in the total amount of lipids in the cells grown in the presence of 8% (vol/vol) ethanol. Preexposure for 2 h to 12% (vol/vol) ethanol also reduced membrane fluidity and cF leakage. This short-term adaptation was not prevented in the presence of chloramphenicol, suggesting that de novo protein synthesis was not involved. We found a strong correlation between fluidity and cF leakage for all treatments and alcohol concentrations tested. We propose that the protective effect of growth in the presence of ethanol is, to a large extent, based on modification of the physicochemical state of the membrane, i.e., cells adjust their membrane permeability by decreasing fluidity at the lipid-water interface.
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flow cytometric assessment of membrane integrity of ethanol stressed Oenococcus oeni cells
Applied and Environmental Microbiology, 2002Co-Authors: Graca M Da Silveira, F M Rombouts, Vitoria San M Romao, Maria C Loureirodias, Tjakko AbeeAbstract:The practical application of commercial malolactic starter cultures of Oenococcus oeni surviving direct inoculation in wine requires insight into the mechanisms involved in ethanol toxicity and tolerance in this organism. Exposure to ethanol resulted in an increase in the permeability of the cytoplasmic membrane, enhancing passive proton influx and concomitant loss of intracellular material (absorbing at 260 nm). Cells grown in the presence of 8% (vol/vol) ethanol revealed adaptation to ethanol stress, since these cells showed higher retention of compounds absorbing at 260 nm. Moreover, for concentrations higher than 10% (vol/vol), lower rates of passive proton influx were observed in these ethanol-adapted cells, especially at pH 3.5. The effect of ethanol on O. oeni cells was studied as the ability to efficiently retain carboxyfluorescein (cF) as an indicator of membrane integrity and enzyme activity and the uptake of propidium iodide (PI) to assess membrane damage. Flow cytometric analysis of both ethanol-adapted and nonadapted cells with a mixture of the two fluorescent dyes, cF and PI, revealed three main subpopulations of cells: cF-stained intact cells; cF- and PI-stained permeable cells, and PI-stained damaged cells. The subpopulation of O. oeni cells that maintained their membrane integrity, i.e., cells stained only with cF, was three times larger in the population grown in the presence of ethanol, reflecting the protective effect of ethanol adaptation. This information is of major importance in studies of microbial fermentations in order to assign bulk activities measured by classical methods to the very active cells that are effectively responsible for the observations.
Aline Lonvaud-funel - One of the best experts on this subject based on the ideXlab platform.
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Probiotic properties of non-conventional lactic acid bacteria: Immunomodulation by Oenococcus oeni
International Journal of Food Microbiology, 2010Co-Authors: Benoit Foligne, Aline Lonvaud-funel, Joelle Dewulf, Jérôme Breton, Olivier Claisse, Bruno PotAbstract:The widely used probiotic bacteria belong to the genera Lactobacillus and Bifidobacterium and have in most cases been isolated from the human gastrointestinal tract. However, other “less conventional” bacteria, from allochthonous or extremophilic origin, sharing similar structural or functional features, may also confer specific health benefits to a host. Firstly, we explored the in vitro immuno-modulatory or immune-stimulatory activities of 25 wine lactic acid bacteria belonging to Oenococcus oeni and Pediococcus parvulus. While cytokines released by peripheral blood mononuclear cells (PBMCs) stimulated by P. parvulus strains, showed little variation, O. oeni strains induced strain-specific cytokine patterns. Some O. oeni strains were then further analyzed under various conditions for growth, dose and culture medium. In a second phase, we evaluated the oral tolerance and safety of two strains of O. oeni in mice fed a high dose of bacteria for a week. Finally, evidence was gathered on the in vivo anti-inflammatory potential of a selected O. oeni strain using an experimental 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis mouse model. Although results did not match the anti-inflammatory levels obtained with certain conventional probiotics, strain IOEB 9115 significantly lowered colonic injury and alleviated colitis symptoms. The ‘natural’ tolerance towards acid, ethanol, and phenolic compounds of O. oeni strains combined with a measureable immunomodulatory potential, suggest a possible use of selected strains isolated from wine as live probiotics.
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Evidence of distinct populations and specific subpopulations within the species Oenococcus oeni
Applied and Environmental Microbiology, 2010Co-Authors: Julen Bridier, Olivier Claisse, Monika Coton, Emmanuel Coton, Aline Lonvaud-funelAbstract:Among the lactic acid bacteria (LAB) present in the oenological microbial ecosystem, Oenococcus oeni, an acidophilic lactic acid bacterium, is essential during winemaking. It outclasses all other bacterial species during malolactic fermentation (MLF). Oenological performances, such as malic acid degradation rate and sensorial impact, vary significantly according to the strain. The genetic diversity of the O. oeni species was evaluated using a multilocus sequence typing (MLST) scheme. Seven housekeeping genes were sequenced for a collection of 258 strains that had been isolated all over the world (particularly Burgundy, Champagne, and Aquitaine, France, Chile, South Africa, and Italy) and in several wine types (red wines, white wines, and champagne) and cider. The allelic diversity was high, with an average of 20.7 alleles per locus, many of them being rare alleles. The collection comprised 127 sequence types, suggesting an important genotypic diversity. The neighbor-joining phylogenetic tree constructed from the concatenated sequence of the seven housekeeping genes showed two major phylogenetic groups, named A and B. One unique strain isolated from cider composed a third group, rooting the phylogenetic tree. However, all other strains isolated from cider were in group B. Eight phylogenetic subgroups were statistically differentiated and could be delineated by the analysis of only 32 mutations instead of the 600 mutations observed in the concatenated sequence of the seven housekeeping genes. Interestingly, in group A, several phylogenetic subgroups were composed mostly of strains coming from a precise geographic origin. Three subgroups were identified, composed of strains from Chile, South Africa, and eastern France.
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Evidence for exopolysaccharide production by Oenococcus oeni strains isolated from non-ropy wines
Journal of Applied Microbiology, 2009Co-Authors: G. Ciezack, Aline Lonvaud-funel, L. Hazo, Alain Heyraud, Gérard Chambat, Marguerite Dols-lafargueAbstract:Aims: The aim of this study was to assess the exopolysaccharide (EPS) production capacities of various strains of Oenococcus oeni, including malolactic starters and strains recently isolated from wine. Methods and Results: Fourteen O. oeni strains displaying or not (PCR check on genomic DNA) the gtf gene generally associated with β-glucan formation and ropiness were grown on grape juice medium, dialysed MRS-derived medium or synthetic medium. The soluble polysaccharides (PS) remaining in the culture supernatant were alcohol precipitated, and their concentration was quantified by the phenol-sulfuric method. Most of the O. oeni strains studied produced significant amounts of EPS, independently of their genotype (gtf+ or gtf−). The EPS production was not directly connected with growth and could be stimulated by changing the growth medium composition. The molecular weight distribution analysis and attempts to determine the PS chemical structure suggested that most strains produce a mixture of EPS. Conclusion: Oenococcus oeni strains recently isolated from wine or cultivated for many generations as a malolactic starter are able to produce EPS other than β-glucan. Significance and Impact of the Study: These EPS may enhance the bacteria survival in wine (advantage for malolactic starters) and may contribute to the wine colloidal equilibrium.
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Oenococcus oeni genome plasticity is associated with fitness
Applied and Environmental Microbiology, 2009Co-Authors: Elisabeth Bon, Aline Lonvaud-funel, Arnaud Delaherche, Eric Bilhere, Antoine De Daruvar, Claire Le MarrecAbstract:Oenococcus oeni strains are well-known for their considerable phenotypic variations in terms of tolerance to harsh wine conditions and malolactic activity. Genomic subtractive hybridization (SH) between two isolates with differing enological potentials was used to elucidate the genetic bases of this intraspecies diversity and identify novel genes involved in adaptation to wine. SH revealed 182 tester-specific fragments corresponding to 126 open reading frames (ORFs). A large proportion of the chromosome-related ORFs resembled genes involved in carbohydrate transport and metabolism, cell wall/membrane/envelope biogenesis, and replication, recombination, and repair. Six regions of genomic plasticity were identified, and their analysis suggested that both limited recombination and insertion/deletion events contributed to the vast genomic diversity observed in O. oeni. The association of selected sequences with adaptation to wine was further assessed by screening a large collection of strains using PCR. No sequences were found to be specific to highly performing (HP) strains alone. However, there was a statistically significant positive association between HP strains and the presence of eight gene sequences located on regions 2, 4, and 5. Gene expression patterns were significantly modified in HP strains, following exposure to one or more of the common stresses in wines. Regions 2 and 5 showed no traces of mobile elements and had normal GC content. In contrast, region 4 had the typical hallmarks of horizontal transfer, suggesting that the strategy of acquiring genes from other bacteria enhances the fitness of O. oeni strains.
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Correlation between indigenous Oenococcus oeni strain resistance and the presence of genetic markers
Journal of Industrial Microbiology and Biotechnology, 2008Co-Authors: Vincent Renouf, Olivier Claisse, Arnaud Delaherche, Aline Lonvaud-funelAbstract:This study reports on monitoring Oenococcus oeni intraspecific diversity evolution during winemaking. Three different wines were monitored. The proportion of O. oeni species was determined by species-specific PCR and O. oeni strains were distinguished by multiplex PCR-RAPD. Each strain was tested by PCR for 16 significant markers revealed by a previous genetic comparison between a strong oenological potential strain and one with poor oenological potential. Population levels and diversity changed according to winemaking stages, oenological practices and the chemical properties of the wine. In all situations, O. oeni was the best-adapted species. Within the O. oeni group, intraspecific strain diversity decreased and the malolactic fermentation was the result of the most resistant strains with the highest number of markers.
Olivier Claisse - One of the best experts on this subject based on the ideXlab platform.
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Characterization of the first virulent phage infecting Oenococcus oeni, the queen of the cellars
Frontiers in Microbiology, 2021Co-Authors: Cécile Philippe, Olivier Claisse, Fety Jaomanjaka, Patrick Lucas, Amel Chaib, Johan Samot, Christian Cambillau, Claire Le MarrecAbstract:There has been little exploration of how phages contribute to the diversity of the bacterial community associated with winemaking and may impact fermentations and product quality. Prophages of Oenococcus oeni, the most common species of lactic acid bacteria (LAB) associated with malolactic fermentation of wine, have been described, but no data is available regarding phages of O. oeni with true virulent lifestyles. The current study reports on the incidence and characterization of the first group of virulent oenophages named Vinitor, isolated from the enological environment. Vinitor phages are morphologically very similar to siphoviruses infecting other LAB. Although widespread during winemaking, they are more abundant in musts than temperate oenophages. We obtained the complete genomic sequences of phages Vinitor162 and Vinitor27, isolated from white and red wines, respectively. The assembled genomes shared 97.6% nucleotide identity and belong to the same species. Coupled with phylogenetic analysis, our study revealed that the genomes of Vinitor phages are architecturally mosaics and represent unique combinations of modules amongst LAB infecting-phages. Our data also provide some clues to possible evolutionary connections between Vinitor and (pro)phages associated to epiphytic and insect-related bacteria.
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Complete genome sequence of lytic Oenococcus oeni bacteriophage OE33PA
Microbiology Resource Announcements, 2018Co-Authors: Fety Jaomanjaka, Olivier Claisse, Cécile Philippe, Claire Le MarrecAbstract:Oenococcus oeni is the most common species of lactic acid bacteria associated with malolactic fermentation in wine. Here, we report the genome sequence of the lytic phage OE33PA (vB_OeS_OE33PA). It has a morphotype similar to that of members of the Siphoviridae family, a linear 39,866-bp double-stranded genome with cohesive ends, and 57 predicted open reading frames.
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Oenococcus oeni Exopolysaccharide Biosynthesis, a Tool to Improve Malolactic Starter Performance
Frontiers Media S.A., 2018Co-Authors: Maria Dimopoulou, Olivier Claisse, Cecile Miot-sertier, Jerôme Raffenne, Nerea Iturmendi, Virginie Moine, Joana Coulon, Marguerite Dols-lafargueAbstract:Oenococcus oeni is the lactic acid bacterium that most commonly drives malolactic fermentation (MLF) in wine. Though the importance of MLF in terms of wine microbial stability and sensory improvement is well established, it remains a winemaking step not so easy to control. O. oeni displays many adaptation tools to resist the harsh wine conditions which explain its natural dominance at this stage of winemaking. Previous findings showed that capsular polysaccharides and endogenous produced dextran increased the survival rate and the conservation time of malolactic starters. In this paper, we showed that exopolysaccharides specific production rates were increased in the presence of single stressors relevant to wine (pH, ethanol). The transcription of the associated genes was investigated in distinct O. oeni strains. The conditions in which eps genes and EPS synthesis were most stimulated were then evaluated for the production of freeze dried malolactic starters, for acclimation procedures and for MLF efficiency. Sensory analysis tests on the resulting wines were finally performed
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Exopolysaccharides produced by Oenococcus oeni: from genomic and phenotypic analysis to technological valorization
Food Microbiology, 2016Co-Authors: Maria Dimopoulou, Olivier Claisse, Tiphaine Bardeau, Pierre-yves Ramonet, Cecile Miot-sertier, Thierry Doco, Melina Petrel, Patrick Lucas, Marguerite Dols-lafargueAbstract:Oenococcus oeni (O. oeni), which is the main species that drives malolactic fermentation (FML), an essential step for wine microbial stabilization and quality improvement, is known to produce exopolysaccharides (EPS). Depending on the strain, these EPS can be soluble, remain attached to the cell or both. In the present study, fourteen strains were examined for eps gene content and EPS production capacities. Cell-linked and soluble heteropolysaccharides made of glucose, galactose and rhamnose, soluble β-glucan, and soluble dextran or levan were found, depending on the strain. The protective potential of either cell-linked heteropolysaccharides or dextrans produced was then studied during freeze drying of the bacterial strains.
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Characterization of a new virulent phage infecting the lactic acid bacterium Oenococcus oeni
Food Microbiology, 2016Co-Authors: Fety Jaomanjaka, Olivier Claisse, Melina Petrel, Mélanie Blanche-barbat, Patricia Ballestra, Claire Le MarrecAbstract:Phi OE33PA is a new virulent siphophage infecting Oenococcus oeni that was isolated from a red wine collected in Pauillac (France). Although the phage could not lysogenize its host, a conserved sequence within the integrase genes harbored by oenophages could be detected, and corresponded to a B-type integrase. The phage host range encompassed ten out of the 38 O. oeni strains tested. One-step growth kinetics revealed latent and burst periods of 4 and 5 h, respectively, with a burst size of about 45 plaque-forming units per infected cell. The phage had a distinctive restriction profile when compared with Phi 10 MC, another B-type oenophage previously isolated in our laboratory. Incubation in wine could inactivate high-titer suspensions of FOE33PA in a short time at room temperature. However, encapsidated phage DNA could still be detected by real time PCR, and these non-infectious viruses dominated the wine samples showing that direct enumeration of phages in wine samples using the double-layer agar technique only informs about the quantity of residual infectious phages. Kinetics studies throughout the fermentation using both qPCR as well as plating should now provide reliable understanding of the phage dynamics during wine making.
Cristina Reguant - One of the best experts on this subject based on the ideXlab platform.
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non saccharomyces in wine effect upon Oenococcus oeni and malolactic fermentation
Frontiers in Microbiology, 2018Co-Authors: Aitor Balmaseda, Albert Bordons, Cristina Reguant, Joaquin BautistagallegoAbstract:This work is a short review of the interactions between oenological yeasts and lactic acid bacteria (LAB), especially Oenococcus oeni, the main species carrying out the malolactic fermentation (MLF). The emphasis has been placed on non-Saccharomyces effects due to their recent increased interest in winemaking. Those interactions are variable, ranging from inhibitory, to neutral and stimulatory and are mediated by some known compounds, which will be discussed. One phenomena responsible of inhibitory interactions is the media exhaustion by yeasts, and particularly a decrease in L-malic acid by some non-Saccharomyces. Clearly ethanol is the main inhibitory compound of LAB produced by S. cerevisiae, but non-Saccharomyces can be used to decrease it. Sulfur dioxide and medium chain fatty acids (MCFAs) produced by yeasts can exhibit inhibitory effect upon LAB or even result lethal. Interestingly mixed fermentations with non-Saccharomyces present less MCFA concentration. Among organic acids derived as result of yeast metabolism, succinic acid seems to be the most related with MLF inhibition. Several protein factors produced by S. cerevisiae inhibiting O. oeni have been described, but they have not been studied in non-Saccharomyces. According to the stimulatory effects, the use of non-Saccharomyces can increase the concentration of favorable mediators such as citric acid, pyruvic acid, or other compounds derived of yeast autolysis such as peptides, glucans, or mannoproteins. The emergence of non-Saccharomyces in winemaking present a new scenario in which MLF has to take place. For this reason, new tools and approaches should be explored to better understand this new winemaking context.
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Selection and characterization of autochthonous strains of Oenococcus oeni for vinification in Priorat (Catalonia, Spain)
OENO One, 2018Co-Authors: Judit Franquès, Patrick M. Lucas, Cristina Reguant, Isabel Araque, Mariette El Khoury, Albert BordonsAbstract:The use of autochthonous strains of Oenococcus oeni for inducing malolactic fermentation (MLF) in wines is increasing because they are well adapted to the conditions of a specific area. The main aim of this work was to select O. oeni strains from Priorat (Catalonia, Spain) wines that would be able to carry out the MLF while maintaining the characteristics of the wine. Forty-five autochthonous strains were selected based on their degradation of L-malic acid, resistance to low pH and high ethanol, and the absence of biogenic amine genes. The three strains with the best characteristics were inoculated and MLF was carried out successfully with the final wines showing good characteristics. One of the inoculated O. oeni strains would be a good candidate to study as possible starter culture. It shows that autochthonous O. oeni strains, once selected, have the potential to be used as an inoculum for wines.
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potential formation of ethyl carbamate in simulated wine inoculated with Oenococcus oeni and lactobacillus plantarum
International Journal of Food Science and Technology, 2009Co-Authors: Silvana V Romero, Albert Bordons, Cristina Reguant, Carme M MasqueAbstract:Summary The production of ethyl carbamate (EC) and its precursor citrulline by two strains of Oenococcus oeni and one of Lactobacillus plantarum during malolactic fermentation (MLF) was studied in different conditions of pH, temperature, and ethanol and l-malic acid concentrations. The potential EC, defined as the concentration of EC after wine is heated at 80 °C for 48 h, was also investigated. The O. oeni strains were able to degrade arginine in the conditions studied and to excrete some citrulline. In these strains, the conditions that led to a slight increase in EC were a high ethanol concentration, low pH and a high l-malic acid concentration. However, the potential EC increased with higher pH. In the case of L. plantarum, arginine was not degraded and citrulline was not produced, although the potential EC was comparable with that of the O. oeni strains studied.
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influence of ethanol and ph on the gene expression of the citrate pathway in Oenococcus oeni
Food Microbiology, 2009Co-Authors: Nair Olguin, Albert Bordons, Cristina ReguantAbstract:The consumption of citrate by the malolactic bacterium Oenococcus oeni changes the aromatic profile of wines due to the production of volatile compounds such as diacetyl and acetic acid. In this study, the expression of genes related to citrate utilization in the O. oeni strain PSU-1 was investigated to further understand the role of this metabolic pathway in the adaptation to wine environment and its impact on organoleptic qualities. Different conditions of ethanol content (0% and 10%) and pH (3.5 and 4.0) were assayed to evaluate the transcriptional response to both these stress factors. In the presence of ethanol, metabolic and transcriptional behavior was different than the observed when ethanol was absent. The expression of citrate pathway genes was mainly affected by ethanol, while pH showed a lower effect. Among the studied genes, citE, ackA and alsD were the genes revealing a distinctive transcriptional response. The differences observed in gene expression were in correlation with the different content of end products such as acetic acid and diacetyl. The increment of gene expression observed in the presence of ethanol at low pH suggests the participation of citrate metabolism in the response to stress conditions.
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genomic dna fingerprinting of Oenococcus oeni strains by pulsed field gel electrophoresis and randomly amplified polymorphic dna pcr
Current Microbiology, 2000Co-Authors: Giacomo Zapparoli, Albert Bordons, Sandra Torriani, Cristina Reguant, Franco DellaglioAbstract:Genetic diversity of 60 Oenococcus oeni strains from different wines was evaluated by numerical analysis of (i) pulsed-field gel electrophoresis (PFGE) patterns with endonuclease ApaI and (ii) randomly amplified polymorphic DNA (RAPD)-PCR fingerprints with four oligonucleotide primers. Sixty-two percent of the strains could be distinguished by PFGE, whereas most strains were identified by distinct RAPD-PCR profiles and associated according to the geographical origin. Because of its rapidity and reliability, RAPD-PCR appeared to be a suitable method for typing and monitoring O. oeni strains in winemaking.