terclim by ICS banner
IVES 9 IVES Conference Series 9 CHANGES IN METABOLIC FLUXES UNDER LOW PH GROWTH CONDITIONS: CAN THE SLOWDOWN OF CITRATE CONSUMPTION IMPROVE OENOCOCCUS OENI ACID-TOLERANCE?

CHANGES IN METABOLIC FLUXES UNDER LOW PH GROWTH CONDITIONS: CAN THE SLOWDOWN OF CITRATE CONSUMPTION IMPROVE OENOCOCCUS OENI ACID-TOLERANCE?

Abstract

Oenococcus oeni is the main Lactic Acid Bacteria responsible for malolactic fermentation, converting malic acid into lactic acid and carbon dioxide in wines. Following the alcoholic fermentation, this second fermentation ensures a deacidification and remains essential for the release of aromatic notes and the improvement of microbial stability in many wines. Nevertheless, wine is a harsh environment for microbial growth, especially because of its low pH (between 2.9 and 3.6 depending on the type of wine) and nutrient deficiency. In order to maintain homeostasis and ensure viability, O. oeni possesses different cellular mechanisms including organic acid metabolisms which represent also the major pathway to synthetize energy in wine. Indeed, uptake and consumption of malate and citrate by this bacteria enables to activate a proton motive force (PMF) hence maintaining an intracellular pH by proton consumption1,2.

Citrate is found in wine at small concentrations (0.13 to 0.90 g/L). It can be metabolized by O. oeni into acetate, pyruvate and then aromatic compounds such as diacetyl, acetoin and 2,3-butanediol. The ability of citrate metabolism to activate the PMF could play a central role in the acid-tolerance of this bacterium. Nevertheless, a previous study has described an inhibition of O. oeni growth at low pH in presence of high amounts of citrate3. This toxic effect could come from the synthesis of one of the citrate metabolites as acetate.

In order to understand how citrate metabolism can be linked to acid tolerance of this bacterium, consumption of citrate was investigated in a great diversity of O. oeni strains. In addition, malate and sugar consumptions were also followed, as they can be impacted by citrate metabolism. These experiments enabled to draw metabolic fluxes in O. oeni according to the pH of the medium. In most cases, ma- late is first metabolized, then citrate and sugars, sequentially, proving that the priority is given to organic acid consumption at the expense of sugars in this bacterium. However, this experiment revealed different citrate consumption profiles which may be correlated to a greater or lesser acid tolerance according to the strain. Furthermore, a genomic comparison demonstrated the presence of mutations in the citrate operon of acid-tolerant industrial strains. Hence, acid tolerance could be linked to a change in the rate of citrate consumption in O. oeni.

 

1. Ramos, A., Poolman, B., Santos, H., Lolkema, J.S., Konings, W.N., 1994. Uniport of anionic citrate and proton consumption in ci-trate metabolism generates a proton motive force in Leuconostoc oenos. J. Bacteriol. 176, 4899–4905. https://doi.org/10.1128/jb.176.16.4899-4905.1994
2. Salema, M., Lolkema, J.S., Romão, M.V.S., Dias, M.C.L., 1996. The proton motive force generated in Leuconostoc oenos by L-malate fermentation. J. Bacteriol. 178, 3127–3132. https://doi.org/10.1128/jb.178.11.3127-3132.1996
3. Augagneur, Y., Ritt, J.-F., Linares, D.M., Remize, F., Tourdot-Maréchal, R., Garmyn, D., Guzzo, J., 2007. Dual effect of organic acids as a function of external pH in Oenococcus oeni. Arch. Microbiol. 188, 147–157. https://doi.org/10.1007/s00203-007-0230-0

DOI:

Publication date: February 9, 2024

Issue: OENO Macrowine 2023

Type: Article

Authors

Camille Eicher1, Joana Coulon2, Marion Favier2, Edouard Munier1, Thierry Tran1, Hervé Alexandre1, Cristina Reguant Miran-da3, Cosette Grandvalet1

1. UMR A. 02.102 Procédés Alimentaires et Microbiologiques, L’institut Agro Dijon, Université de Bourgogne Franche-Comté, Dijon, France
2. BioLaffort, Floirac, France
3. Departament de Bioquímica i Biotecnologia, Facultat d’Enologia, Universitat Rovira i Virgili, Tarragona, Spain

Contact the author*

Keywords

Oenococcus oeni, Citrate, Metabolic fluxes, Acid-tolerance

Tags

IVES Conference Series | oeno macrowine 2023 | oeno-macrowine

Citation

Related articles…

ACIDIC AND DEMALIC SACCHAROMYCES CEREVISIAE STRAINS FOR MANAGING PROBLEMS OF ACIDITY DURING THE ALCOHOLIC FERMENTATION

In a recent study several genes controlling the acidification properties of the wine yeast Saccharomyces cerevisiae have been identified by a QTL approach [1]. Many of these genes showed allelic variations that affect the metabolism of malic acid and the pH homeostasis during the alcoholic fermentation. Such alleles have been used for driving genetic selection of new S. cerevisiae starters that may conversely acidify or deacidify the wine by producing or consuming large amount of malic acid [2]. This particular feature drastically modulates the final pH of wine with difference of 0.5 units between the two groups.

DOES LIGNIN AN ACCEPTABLE MARKER OF GRAPESEED MATURATION AND QUALITY?

Usually the winemaker consider polyphenols from the grape berry as an actor of the wine quality. There are frequently consider as a marker of grape maturity. It is commonly known that winemaker consider tannins and anthocyanins as main polyphenol actors for winemaking practices and wine quality. Here we will focus on the characterisation of lignins in grape seeds. Previous studies suggest that the seed is lignified [1], which could explain the change in colour of the seed when it reaches maturity and thus provide a reliable indicator for describing the maturity stage in the seed.

METHYL SALICYLATE, A COMPOUND INVOLVED IN BORDEAUX RED WINES PRODUCED WITHOUT SULFITES ADDITION

Sulfur dioxide (SO₂) is the most commonly used additive during winemaking to protect wine from oxidation and from microorganisms. Thus, since the 18th century, SO₂ was almost systematically present in wines. Recently, wines produced without any addition of SO₂ during all the winemaking process including bottling became more and more popular for consumers. A recent study dedicated to sensory characterization of Bordeaux red wines produced without added SO₂, revealed that such wines were perceived differently from similar wines produced with using SO₂ and were characterized by specific fruity aromas and coolness1,2.

IMPACT OF RHIZOPUS AND BOTRYTIS ON WINE FOAMING PROPERTIES

A lot of work has been done on the impact of Botrytis on the foam of sparkling wines. This work often concerns wines produced in cool regions, where Botrytis is the dominant fungal pathogen. However, in southern countries such as Spain, in particularly hot years such as 2022, the majority fungal pathogen is sometimes Rhizopus. Like Botrytis, Rhizopus is a fungus that produces an aspartic protease.

INOCULATION OF THE SELECTED METSCHNIKOWIA PULCHERRIMA MP1 AS A BIOPROTECTIVE ALTERNATIVE TO SULFITES TO PREVENT BROWNING OF WHITE GRAPE MUST

Enzymatic browning (BE) of must is caused by polyphenol oxidases (PPOs), tyrosinase and laccase. Both PPOs can oxidize diphenols such as hydroxycinnamic acids (HA) to quinones, which can later polymerize to form melanins [1], which are responsible of BE in white wines and of oxidasic haze in red wines. SO₂ is the main tool used to protect must from BE thanks to its capacity to inhibit PPOs [2]. However, the current trend in winemaking is to reduce and even eliminate this unfriendly additive. Among the different possible alternatives for protecting must against BE, the inoculation with a selected Metschnikowia pulcherrima MP1 is without any doubt one of the most promising ones.