Wine Bacteria Research Articles

Citrate metabolism in lactic acid bacteria: is there a beneficial effect for Oenococcus oeni in wine?

Lactic acid bacteria (LAB) are Gram positive bacteria frequently used in the food industry for fermentation, mainly transformation of carbohydrates into lactic acid. In addition, these bacteria also have the capacity to metabolize citrate, an organic acid commonly found in food products. Its fermentation leads to the production of 4-carbon compounds such as diacetyl, resulting in a buttery flavor desired in dairy products. Citrate metabolism is known to have several beneficial effects on LAB physiology. Nevertheless, a controversial effect of citrate has been described on the acid tolerance of the wine bacterium Oenococcus oeni. This observation raises questions about the effect of citrate on the capacity of O. oeni to conduct malolactic fermentation in highly acidic wines. This review aims to summarize the current understanding of citrate metabolism in LAB, with a focus on the wine bacterium O. oeni. Metabolism with the related enzymes is detailed, as are the involved genes organized in cit loci. The known systems of cit locus expression regulation are also described. Finally, the beneficial effects of citrate catabolism on LAB physiology are reported and the negative impact observed in O. oeni is discussed.

Bacterial community structure of rice wine from China's Xiaogan and Dazhou regions: Correlation with taste characteristics

To investigate the bacterial profiles and taste characteristics of rice wine from the Xiaogan and Dazhou regions, this study employed a combination of High-throughput sequencing, traditional pure culture techniques, and bionics equipment. The findings revealed a significant difference (P < 0.05) in bacterial diversity and community structure between the rice wine from Xiaogan and Dazhou. In particular, Leuconostoc, Lactococcus, Lacticaseibacillus, Exiguobacterium, Gluconobacter, Streptococcus, and Pantoea were significantly enriched in Xiaogan rice wine, Limosilactobacillus, Pediococcus, and Lactobacillus were significantly enriched in Dazhou rice wine. Notably, the most dominant group of bacteria in rice wine was lactic acid bacteria (LAB), and the main culturable isolates were Limosilactobacillus fermentum and Pediococcus pentosaceus. Additionally, the dominant bacterial genera showed a significant correlation with the taste characteristics of rice wine. Limosilactobacillus, Pediococcus, Lactobacillus, and Pseudomonas were positively correlated with the lactic acid, citric acid, pH value, Ala, Ile, and Leu of rice wine. Meanwhile, Exiguobacterium, Lacticaseibacillus, and Streptococcus were positively correlated with the richness of rice wine. This study provides insights into the bacterial diversity and taste characteristics of rice wine from different regions of the Yangtze River Basin and highlights the potential of traditional fermented foods as resources for LAB.

Profiling the Bacterial Community of Fermenting Traminette Grapes during Wine Production using Metagenomic Amplicon Sequencing.

Advances in sequencing technology and the relatively easy access to the use of bioinformatics tools to profile microbial community structures have facilitated a better understanding of both culturable and non-culturable microbes in grapes and wine. During industrial fermentation, microbes, known and unknown, are often responsible for product development and off-flavor. Therefore, profiling the bacteria from grape to wine can enable an easy understanding of in situ microbial dynamics. In this study, the bacteria of Traminette grapes must undergoing fermentation, and the final wine were subjected to DNA extraction that yielded 15 ng/µL to 87 ng/µL. The 16S amplicon of the hypervariable region of the V4 region was sequenced, relatively abundant bacteria consisting of phyla Proteobacteria, Actinobacteriota, Firmicutes, Bacteroidota, Fusobacteriota and followed by the Verrucomicrobiota, Halobacterota, Desulfobacterota, Myxococcota, and Acidobacteriota. A Venn diagram analysis of the shared unique operational taxonomic units (OTU) revealed that 15 bacteria phyla were common to both grape must, fermenting stage, and final wine. Phyla that were not previously reported were detected using the 16S amplicon sequencing, as well as genera such as Enterobacteriaceae and Lactobacillaceae. Variation in the organic nutrient use in wine and its impact on bacteria was tested; Traminette R tank containing Fermaid O and Traminette L stimulated with Stimula Sauvignon blanc + Fermaid O. Alpha diversity using the Kruskal-Wallis test determined the degree of evenness. The beta diversity indicated a shift in the bacteria at the fermentation stage for the two treatments, and the final wine bacteria looked similar. The study confirmed that 16S amplicon sequencing can be used to monitor bacteria changes during wine production to support quality and better utilization of grape bacteria during wine production.

Development and Validation of a New Method for Detecting Acetic Bacteria in Wine.

In winemaking, excessive production of acetic acid by acetic acid bacteria poses a major challenge, leading to rejection of wine by consumers. The aim of this study was to devise an economically viable and easy-to-use liquid culture medium for the preventive detection of microorganisms capable of generating acetic acid in wine. The modified medium incorporated specific nutrients that favored the growth of acetic acid bacteria and increased selectivity. Under varying conditions and with different types of wine, this medium was tested together with inoculated samples, comparing the occurrence of acetic acid and olfaction. The result was a new liquid medium based on olfactometry, designed to facilitate its use in wineries, even by untrained personnel and without the need for complex laboratory equipment. Validation was carried out on a variety of wines, determining the onset of the presence of acetic acid in the medium. This innovative culture medium provides a means to estimate the concentration of micro-organisms capable of producing acetic acid in wine. Its application in wineries facilitates proactive decision making, avoiding undesirable increases in acetic acid concentration.

Characterization of tongue dorsum microbiome in wine tasters

Taste plays a paramount role in food and beverage choice, with recent studies pointing to a potential influence of the microorganisms from the tongue dorsum - particularly bacteria - on flavor perception. Thus, the association between tongue dorsum biofilm and taste is a fundamental prerequisite for a better understanding of the role played by these bacteria in wine tasting. To study this impact, we have analyzed the microbiomes from 58 samples of the tongue dorsum surface from professional wine tasters and 30 samples from non professional wine tasters. The microbiome of each sample was characterized through metagenome sequencing of the 16S rRNA gene for taxonomic discrimination of bacteria. A total of 497 taxa were identified in the tongue dorsum, and significant differences in diversity were observed between the wine taster and the control group. The comparison of bacterial diversity between samples collected before and after wine tasting along with the presence of new bacterial taxa indicates a direct effect of wine on the microbiome of frequent wine tasters, particularly in those tasting sparkling wines.

Exploring the diversity of bacteriophage specific to Oenococcus oeni and Lactobacillus spp and their role in wine production.

The widespread existence of bacteriophage has been of great interest to the biological research community and ongoing investigations continue to explore their diversity and role. They have also attracted attention and in-depth research in connection to fermented food processing, in particular from the dairy and wine industries. Bacteriophage, mostly oenophage, may in fact be a 'double edged sword' for winemakers: whilst they have been implicated as a causal agent of difficulties with malolactic fermentation (although not proven), they are also beginning to be considered as alternatives to using sulphur dioxide to prevent wine spoilage. Investigation and characterisation of oenophage of Oenococcus oeni, the main species used in winemaking, are still limited compared to lactococcal bacteriophage of Lactococcus lactis and Lactiplantibacillus plantarum (formally Lactobacillus plantarum), the drivers of most fermented dairy products. Interestingly, these strains are also being used or considered for use in winemaking. In this review, the genetic diversity and life cycle of phage, together with the debate on the consequent impact of phage predation in wine, and potential control strategies are discussed. KEY POINTS: • Bacteriophage detected in wine are diverse. • Many lysogenic bacteriophage are found in wine bacteria. • Phage impact on winemaking can depend on the stage of the winemaking process. • Bacteriophage as potential antimicrobial agents against spoilage organisms.

Structure and Topology Prediction of Phage Adhesion Devices Using AlphaFold2: The Case of Two Oenococcus oeni Phages.

Lactic acid bacteria (LAB) are important microorganisms in food fermentation. In the food industry, bacteriophages (phages or bacterial viruses) may cause the disruption of LAB-dependent processes with product inconsistencies and economic losses. LAB phages use diverse adhesion devices to infect their host, yet the overall picture of host-binding mechanisms remains incomplete. Here, we aimed to determine the structure and topology of the adhesion devices of two lytic siphophages, OE33PA and Vinitor162, infecting the wine bacteria Oenococcus oeni. These phages possess adhesion devices with a distinct composition and morphology and likely use different infection mechanisms. We primarily used AlphaFold2, an algorithm that can predict protein structure with unprecedented accuracy, to obtain a 3D model of the adhesion devices’ components. Using our prior knowledge of the architecture of the LAB phage host-binding machineries, we also reconstituted the topology of OE33PA and Vinitor162 adhesion devices. While OE33PA exhibits original structures in the assembly of its bulky adhesion device, Vinitor162 harbors several carbohydrate-binding modules throughout its long and extended adhesion device. Overall, these results highlight the ability of AlphaFold2 to predict protein structures and illustrate its great potential in the study of phage structures and host-binding mechanisms.

Analysis of Transcriptomic Response to SO2 by Oenococcus oeni Growing in Continuous Culture.

ABSTRACTTo successfully complete malolactic fermentation (MLF), Oenococcus oeni must overcome wine stress conditions of low pH, high ethanol, and the presence of SO2. Failure to complete MLF may result in detrimental effects to the quality and stability of the resulting wines. Research efforts to date have focused on elucidating the mechanisms and genetic features that confer the ability to withstand low pH and high ethanol concentrations on O. oeni; however, the responses to SO2 stress are less well defined. This study focused on characterizing the transcriptional response of O. oeni to SO2 challenge during cultivation in a continuous system at wine-like pH (3.5). This experimental design allowed the precise discrimination of transcriptional changes linked to SO2 stress from responses associated with growth stage and cultivation parameters. Differential gene expression analysis revealed major transcriptional changes following SO2 exposure and suggested that this compound primarily interacts with intracellular proteins, DNA, and the cell envelope of O. oeni. The molecular chaperone hsp20, which has a demonstrated function in the heat, ethanol, and acid stress response, was highly upregulated, confirming its additional role in the response of this species to SO2 stress. This work also reports the first nanopore-based complete genome assemblies for O. oeni.IMPORTANCE Malolactic fermentation is an indispensable step in the elaboration of most wines and is generally performed by Oenococcus oeni, a Gram-positive heterofermentative lactic acid bacterium species. While O. oeni is tolerant to many of the wine stresses, including low pH and high ethanol concentrations, it has high sensitivity to SO2, an antiseptic and antioxidant compound regularly used in winemaking. Understanding the physiological changes induced in O. oeni by SO2 stress is essential for the development of more robust starter cultures and methods for their use. This study describes the main transcriptional changes induced by SO2 stress in the wine bacterium O. oeni and provides foundational understanding on how this compound interacts with the cellular components and the induced protective mechanisms of this species.

Exopolysaccharides Producing Lactic Acid Bacteria in Wine and Other Fermented Beverages: For Better or for Worse?

Lactic acid bacteria (LAB) from fermented beverages such as wine, cider and beer produce a wide range of exopolysaccharides (EPS) through multiple biosynthetic pathways. These extracellular polysaccharides constitute key elements for bacterial species adaptation to such anthropic processes. In the food industry, LAB polysaccharides have been widely studied for their rheological, functional and nutritional properties; however, these have been poorly studied in wine, beer and cider until recently. In this review, we have gathered the information available on these specific polysaccharide structure and, biosynthetic pathways, as well as the physiology of their production. The genes associated with EPS synthesis are also presented and compared. Finally, the possible role of EPS for bacterial survival and spread, as well as the risks or possible benefits for the winemaker and the wine lover, are discussed.

Microbial interactions in alcoholic beverages

This review examines the different types of interactions between the microorganisms involved in the fermentation processes of alcoholic beverages produced all over the world from cereals or fruit juices. The alcoholic fermentation converting sugars into ethanol is usually carried out by yeasts, mainly Saccharomyces cerevisiae, which can grow directly using fruit sugars, such as those in grapes for wine or apples for cider, or on previously hydrolyzed starch of cereals, such as for beers. Some of these beverages, or the worts obtained from cereals, can be distilled to obtain spirits. Besides S. cerevisiae, all alcoholic beverages can contain other microorganisms and especially in spontaneous fermentation when starter cultures are not used. These other microbes are mostly lactic acid bacteria and other yeasts-the non-Saccharomyces yeasts. The interactions between all these microorganisms are very diverse and complex, as in any natural occurring ecosystem, including food fermentations. To describe them, we have followed a simplified ecological classification of the interactions. The negative ones are amensalism, by which a metabolic product of one species has a negative effect on others, and antagonism, by which one microbe competes directly with others. The positive interactions are commensalism, by which one species has benefits but no apparent effect on others, and synergism, by which there are benefits for all the microbes and also for the final product. The main interactions in alcoholic beverages are between S. cerevisiae and non-Saccharomyces and between yeasts and lactic acid bacteria. These interactions can be related to metabolites produced by fermentation such as ethanol, or to secondary metabolites such as proteinaceous toxins, or are feed-related, either by competition for nutrients or by benefit from released compounds during yeast autolysis. The positive or negative effects of these interactions on the organoleptic qualities of the final product are also revised. Focusing mainly on the alcoholic beverages produced by spontaneous fermentations, this paper reviews the interactions between the different yeasts and lactic acid bacteria in wine, cider, beer, and in spirits such as tequila, mezcal and cachaça.

Relationship between lactic acid bacteria, malolactic fermentation and wine colour

Some phenolic acids can either inhibit or stimulate the growth of Oenococcus oeni and other lactic acid bacteria (LAB) in wine. It has been observed that some LAB metabolism could have an influence on wine colour. In this article, some of the relationships between LAB, malolactic fermentation (MLF) and phenolic compounds are summarised; these relationships are important for the selection of LAB to make starter cultures and are of interest for wineries in terms of its effect on wine colour.

Yeast and lactic acid population relationship in palm wine and their effects on the production of ethanol in Enugu Ezike south eastern Nigeria

Palm wine produced traditionally and consumed by many people in the South eastern part of Nigeria is a whitish liquid formed by naturally fermenting the sugary sap from various palm plants. In this study, we evaluated the population of yeast and lactic acid bacteria in palm wine with regards to its ethanol concentration from three communities in Enugu Ezike, Enugu State. We monitored the isolation of microbial population as well as biochemical characteristics of fermenting palm wine samples using culture-dependent and culture-independent methods. The fermentation step was performed within time interval and completed after 48hrs. Its acidity increasedprogressively with the production of lactic and acetic acids by bacteria.During the fermentation, palm wine microbial numbers (log10 CFU/ml) varied between 1.6 x104 to 7.6 x107 for yeast and 4.5 x 104 to 7.8 x 107 for lactic acid bacteria respectively. Similarly, pH and alcohol level of palm wine samples during the fermentation varied between 3.0 to 5.8 and 0.2 to 5.3 respectively. Statistical analysis revealed that there was no significant difference in both yeast and lactic acid bacteria population at 24hrs. At this time, there was an increased metabolic activity, leading to an increase in ethanol production. At 36-48hrs, there was a slight decrease in both the lactic acid bacteria and the yeast, indicating a reduction in source of carbon.The analysis of the yeast and the lactic acid bacterial population revealed different yeast and bacterial population. Bacteria compete with yeast for nutrients during ethanol production process, potentially causing economic losses. Therefore, it is important to have a better understanding of the abundance and the change in population of lactic acid bacteria and yeast throughout the bio-ethanol process in order to achieve design a more efficient relationship in lactic acid, yeast and ethanol production processes.&#x0D; Keywords: Bacteria, ethanol, lactic acid, palm wine, yeast

Lactic Acid Bacteria in Wine: Technological Advances and Evaluation of Their Functional Role.

Currently, the main role of Lactic Acid Bacteria (LAB) in wine is to conduct the malolactic fermentation (MLF). This process can increase wine aroma and mouthfeel, improve microbial stability and reduce the acidity of wine. A growing number of studies support the appreciation that LAB can also significantly, positively and negatively, contribute to the sensorial profile of wine through many different enzymatic pathways. This is achieved either through the synthesis of compounds such as diacetyl and esters or by liberating bound aroma compounds such as glycoside-bound primary aromas and volatile thiols which are odorless in their bound form. LAB can also liberate hydroxycinnamic acids from their tartaric esters and have the potential to break down anthocyanin glucosides, thus impacting wine color. LAB can also produce enzymes with the potential to help in the winemaking process and contribute to stabilizing the final product. For example, LAB exhibit peptidolytic and proteolytic activity that could break down the proteins causing wine haze, potentially reducing the need for bentonite addition. Other potential contributions include pectinolytic activity, which could aid juice clarification and the ability to break down acetaldehyde, even when bound to SO2, reducing the need for SO2 additions during winemaking. Considering all these findings, this review summarizes the novel enzymatic activities of LAB that positively or negatively affect the quality of wine. Inoculation strategies, LAB improvement strategies, their potential to be used as targeted additions, and technological advances involving their use in wine are highlighted along with suggestions for future research.

Evaluating the Cytometric Detection and Enumeration of the Wine Bacterium, Oenococcus oeni.

Flow cytometry is a high-throughput tool for determining microbial abundance in a range of medical, environmental, and food-related samples. For wine, determining the abundance of Saccharomyces cerevisiae is well-defined and reliable. However, for the most common wine bacterium, Oenococcus oeni, using flow cytometry to determine cell concentration poses some challenges. O. oeni most often occurs in doublets or chains of varying lengths that can be greater than seven cells. This wine bacterium is also small, at 0.2-0.6 μm and may exhibit a range of morphologies including binary fission and aggregated complexes. This work demonstrates a straightforward approach to determining the suitability of flow cytometry for the chain-forming bacteria, O. oeni, and considerations when using flow cytometry for the enumeration of small microorganisms (<0.5 μm). © 2020 International Society for Advancement of Cytometry.

Lactobacillus plantarum, a New Biological Tool to Control Malolactic Fermentation: A Review and an Outlook

Malolactic fermentation (MLF) in wine is an important step in the vinification of most red and some white wines, as stands for the biological conversion of l-malic acid into l-lactic acid and carbon dioxide, resulting in a decrease in wine acidity. MLF not only results in a biological deacidification, it can exert a significant impact on the organoleptic qualities of wine. This paper reviews the biodiversity of lactic acid bacteria (LAB) in wine, their origin, and the limiting conditions encountered in wine, which allow only the most adapted species and strains to survive and induce malolactic fermentation. Of all the species of wine LAB, Oenococcus oeni is probably the best adapted to overcome the harsh environmental wine conditions and therefore represents the majority of commercial MLF starter cultures. Wine pH is most challenging, but, as a result of global warming, Lactobacillus sp. is more often reported to predominate and be responsible for spontaneous malolactic fermentation. Some Lactobacillus plantarum strains can tolerate the high alcohol and SO2 levels normally encountered in wine. This paper shows the potential within this species for the application as a starter culture for induction of MLF in juice or wine. Due to its complex metabolism, a range of compositional changes can be induced, which may positively affect the quality of the final product. An example of a recent isolate has shown most interesting results, not only for its capacity to induce MLF after direct inoculation, but also for its positive contribution to the wine quality. Degrading hexose sugars by the homo-fermentative pathway, which poses no risk of acetic acid production from the sugars, is an interesting alternative to control MLF in high pH wines. Within this species, we can expect more strains with interesting enological properties.

Co-Existence of Inoculated Yeast and Lactic Acid Bacteria and Their Impact on the Aroma Profile and Sensory Traits of Tempranillo Red Wine

This study investigates the effects of simultaneous inoculation of a selected Saccharomyces cerevisiae yeast strain with two different commercial strains of wine bacteria Oenococcus oeni at the beginning of the alcoholic fermentation on the kinetics of malolactic fermentation (MLF), wine chemical composition, and organoleptic characteristics in comparison with spontaneous MLF in Tempranillo grape must from Castilla-La Mancha (Spain). Evolution of MLF was assessed by the periodic analysis of L-malic acid through the enzymatic method, and most common physiochemical parameters and sensory traits were evaluated using a standardized sensory analysis. The samples were analyzed by GC/MS in SCAN mode using a Trace GC gas chromatograph and a DSQII quadrupole mass analyzer. Co-inoculation reduced the overall fermentation time by up to 2 weeks leading to a lower increase in volatile acidity. The fermentation-derived wine volatiles profile was distinct between the co-inoculated wines and spontaneous MLF and was influenced by the selected wine bacteria used in co-inoculation. Co-inoculation allows MLF to develop under reductive conditions and results in wines with very few lactic and buttery flavors, which is related to the impact of specific compounds like 2,3-butanedione. This compound has been also confirmed as being dependent on the wine bacteria used.

Bacteriophages as an Up-and-Coming Alternative to the Use of Sulfur Dioxide in Winemaking.

Certain acetic and lactic acid bacteria are major causes of quality defects in musts and wines, giving rise to defects such as a “vinegary,” “sharp, like nail polish-remover” taste or preventing alcoholic and/or malolactic fermentation. Sulfur dioxide is the major tool currently used in the control of these bacteria in wine. The aim of this work was to isolate bacteriophages from musts and wine of different grape varieties that were able to eliminate lactic and acetic acid bacteria spoilages at the laboratory scale. Musts obtained from grape-berries of Vitis vinifera cv. Chardonnay and Moscatel and a red wine made with V. vinifera cv. Tintilla de Rota were used to isolate bacteriophages. Bacteriophages were obtained from each of the musts and the wine and belonged to the order Caudovirals and the family Tectivirals. They were isolated by classical virology methods and identified by electron microscopy. The host bacteria used in the study were lactic acid bacteria of the species Lactobacillus hilgardii, Lactobacillus plantarum, and Oenococcus oeni and the acetic bacteria Acetobacter aceti. A comparative study was performed by adding phage titrations and SO2 to musts and wines, which had been previously inoculated with bacteria, to study the effectiveness of bacteriophages against bacteria. The comparative study showed that some bacteriophages were as effective as sulfur dioxide at low concentrations.

Production and Antimicrobial Activity of Nisin Under Enological Conditions.

Lactic acid bacteria (LAB) are responsible for the malolactic fermentation of wines, and, therefore, controlling the growth of these bacteria is a key factor for elaborating premium wines. Sulfur dioxide has been traditionally used as an efficient antimicrobial and antioxidant agent, however, nowadays consumers’ demand tends toward a reduction of sulfur dioxide levels in wine and other fermented foods. A previous study of our research group had demonstrated the effectiveness of the bacteriocin nisin to inhibit the growth of enological LAB, and its activity had been tested in culture broths. The aim of this study was to investigate the possibility of controlling the growth of bacteria in wine by the use of nisin in combination with sulfur dioxide, and to study nisin production by the natural producer Lactococcus lactis LM29 under enological conditions. Our results showed that L. lactis LM29 produced nisin in the presence of 2 and 4% ethanol (v/v), while higher concentrations of ethanol fully inhibited the production of nisin. We obtained a nisin enriched active extract (NAE) from the cell-free supernatant of a culture of L. lactis LM29 in MRS broth containing 60% (v/v) sterile grape juice, and the extract was fully active in inhibiting the growth of the enological LAB tested by the microtiter method. Moreover, the nisin concentration of the obtained NAE could actually prevent the formation of an undesirable biofilm of LAB strains. Finally, our results of wine ageing under winery conditions showed that the use of 50 mg/L nisin decreased fourfold the concentration of sulfur dioxide required to prevent LAB growth in the wines.

Arginine and Citrulline Catabolic Pathways Encoded by the arc Gene Cluster of Lactobacillus brevis ATCC 367.

High concentrations of l-arginine or l-citrulline in the growth medium provided the wine bacterium Lactobacillus brevis with a significant growth advantage. The arginine deiminase pathway (ADI) arc gene cluster of Lactobacillus brevis contains three genes-arcD, arcE1, and arcE2-encoding putative l-arginine/l-ornithine exchangers. Uptake experiments with Lactococcus lactis cells expressing the genes showed that all three transported l-ornithine with affinities in the micromolar range. Similarly, ArcD and ArcE2 transported l-arginine, while ArcE1 transported l-citrulline, an intermediate of the ADI pathway. Chase experiments showed very efficient exchange of l-arginine and l-ornithine by ArcD and ArcE2 and of l-citrulline and l-ornithine by ArcE1. Low affinities (millimolar range) combined with low translocation rates were found for ArcD and ArcE2 with l-citrulline and for ArcE1 with l-arginine. Resting cells of Lactobacillus brevis grown in the presence of l-arginine and l-citrulline rapidly consumed l-arginine and l-citrulline, respectively, while producing ammonia and l-ornithine. About 10% of l-arginine degraded was excreted by the cells as l-citrulline. Degradation of l-arginine and l-citrulline was not subject to carbon catabolite repression by glucose in the medium. At a high medium pH, l-citrulline in the medium was required for induction of the l-citrulline degradation pathway. Pathways are proposed for the catabolic breakdown of l-arginine and l-citrulline that merge at the level of ornithine transcarbamylase in the ADI pathway. l-Arginine uptake is catalyzed by ArcD and/or ArcE2, l-citrulline by ArcE1. l-Citrulline excretion during l-arginine breakdown is proposed to be catalyzed by ArcD and/or ArcE2 through l-arginine/l-citrulline exchange.IMPORTANCELactobacillus brevis, a bacterium isolated from wine, as well as other food environments, expresses a catabolic pathway for the breakdown of l-citrulline in the medium that consists of the l-citrulline/l-ornithine exchanger ArcE1 and part of the catabolic arginine deiminase (ADI) pathway enzymes. The proposed pathways for l-arginine and l-citrulline breakdown provide a mechanism for l-citrulline accumulation in fermented food products that is the precursor of the carcinogen ethyl carbamate.

Effect of kaolin silver complex on the control of populations of Brettanomyces and acetic acid bacteria in wine.

In this work, the effects of kaolin silver complex (KAgC) have been evaluated to replace the use of SO2 for the control of spoilage microorganisms in the winemaking process. The results showed that KAgC at a dose of 1g/L provided effective control against the development of B. bruxellensis and acetic acid bacteria. In wines artificially contaminated with an initial population of B. bruxellensis at 104 CFU/mL, a concentration proven to produce off flavors in wine, only residual populations of the contaminating yeast remained after 24days of contact with the additive. Populations of acetic bacteria inoculated into wine at concentrations of 102 and 104CFU/mL were reduced to negligible levels after 72h of treatment with KAgC. The antimicrobial effect of KAgC against B. bruxellensis and acetic bacteria was also demonstrated in a wine naturally contaminated by these microorganisms, decreasing their population in a similar way to a chitosan treatment. Related to this effect, wines with KAgC showed lower concentrations of acetic acid and 4-ethyl phenol than wines without KAgC. The silver concentration from KAgC that remained in the finished wines was below the legal limits. These results demonstrated the effectiveness of KAgC to reduce spoilage microorganisms in winemaking.