terclim by ICS banner
IVES 9 IVES Conference Series 9 INVESTIGATION OF MALIC ACID METABOLIC PATHWAYS DURING ALCOHOLIC FERMENTATION USING GC-MS, LC-MS, AND NMR DERIVED 13C-LABELED DATA

INVESTIGATION OF MALIC ACID METABOLIC PATHWAYS DURING ALCOHOLIC FERMENTATION USING GC-MS, LC-MS, AND NMR DERIVED 13C-LABELED DATA

Abstract

Malic acid has a strong impact on wine pH and the contribution of fermenting yeasts to modulate its concentration has been intensively investigated in the past. Recent advances in yeast genetics have shed light on the unexpected property of some strains to produce large amounts of malic acid (“acidic strains”) while most of the wine starters consume it during the alcoholic fermentation. Being a key metabolite of the central carbohydrate metabolism, malic acid participates to TCA and glyoxylate cycles as well as neoglucogenesis. Although present at important concentrations in grape juice, the metabolic fate of malic acid has been poorly investigated. In this work, we used 13C-labeled malic acid to understand the main routes of its consumption and its de novo production. Two strains selected for their opposed malic acid metabolism were compared by combining several analytical chemistry techniques. The isotopic enrichment of intracellular amino acids was measured by GC-MS, the relative quantification of intra- cellular and extracellular labeled compounds was achieved by 2D-NMR, and the absolute quantification of labeled and unlabeled extracellular organic acids was achieved by LC-MS/MS. Although, both strains consume most of the malic acid provided, the “acidic strain” produces de novo malic acid during the second part of the alcoholic fermentation. In addition, 13C-filiation analyses provided evidence that most of the TCA is fed by glycolytic pyruvate and/or by cytosolic acetyl-CoA. Our results also confirmed that malic acid may be a secondary source of TCA cycle during alcoholic fermentation especially in high malic acid consuming strains that has an efficient malo-ethanolic fermentation. Finally, 13C-labeled compounds belonging to amino acids, alcoholic fermentation and neoglucogenesis pathways were identified, highlighting the pleiotropic position of malic acid in both catabolic and anabolic routes.

DOI:

Publication date: February 9, 2024

Issue: OENO Macrowine 2023

Type: Poster

Authors

Vion Charlotte1,2, Bloem Audrey3, Valette Gilles4, Da Costa Gregory2, Richard Tristan2, Camarasa Carole3, Marullo Philippe 1,2

1. Biolaffort, Bordeaux, FRANCE
2. UMR 1366 Œnologie, Université de Bordeaux, INRAE, Bordeaux INP, BSA, ISVV
3. UMR SPO, Univ Montpellier, INRAE, Institut Agro, 34060 Montpellier, France
4. IBMM, Univ Montpellier, CNRS, ENSCM, 34000 Montpellier, France

Contact the author*

Keywords

13C-labeling, malic acid, central carbon metabolism

Tags

IVES Conference Series | oeno macrowine 2023 | oeno-macrowine

Citation

Related articles…

MOUSY OFF-FLAVOURS IN WINES: UNVEILING THE MICROORGANISMS BEHIND IT

Taints and off-flavours are one of the major concerns in the wine industry and even if the issues provoked by them are harmless, they can still have a negative impact on the quality or on the visual perception of the consumer. Nowadays, the frequency of occurrence of mousy off-flavours in wines has increased.
The reasons behind this could be the significant decrease in sulphur dioxide addition during processing, the increase in pH or even the trend for spontaneous fermentation in wine. This off-flavour is associated with Brettanomyces bruxellensis or some lactic acid bacteria metabolisms.

WINE AS AN EMOTIONAL AND AESTHETIC OBJECT: IMPACT OF EXPERTISE

Wine tasting has been shown to provide emotions to tasters (Coste et al. 2018). How will expertise impact this emotional response? Burnham and Skilleås (2012) reported that the cultural, experiential, and aesthetic competencies characterize an expert in wine compared to a novice. Although there is no consensual definition of an aesthetic experience, Burnham and Skilleås (2012) reported that aesthetic appreciation is “disinterested, normative for others and communicable” in comparison to sensory pleasure.

THE EFFECT OF PRE-FERMENTATIVE GLYPHOSATE ADDITION ON THE METABOLITE PROFILE OF WINE

The synthetic herbicide glyphosate has been used extensively in viticulture over many decades to combat weeds. Despite this, the possible influence of residual glyphosate on both the alcoholic fermentation of grape juice and the subsequent metabolite profile of wines has not been investigated. In this study, Pinot noir juice supplemented with different concentrations of glyphosate (0 µg L-1, 10 µg L-1 and 1000 µg L-1) was fermented with commercial Saccharomyces cerevisiae yeast strains. Using a combination of analytical methods, 80 metabolites were quantified in the resulting wines.

IDENTIFICATION AND LEVELS OF PHENOLIC COMPOUNDS (TANINS, ANTHO-CYANS) IN RED VARIETAL WINES (PROKUPAC AND BLACK TAMJANIKA) FROM SERBIA

The phenolic compounds of red wines represent a source of numerous benefits for human health, which is why they are a constant subject of scientific research. Winemaking in Serbia has a growing economic significance, with particularly autochthonous varieties included [1]. This research identifies and quantifies phenolic compounds of Serbian red varietal wines of Prokupac and Black Tamjanika varieties. Quantification of the level of phenolics has been conducted, including molecular tannins [(+)-catechin, (-)-epicatechin, procyanidin dimers B1, B2, B3, B4], molecular anthocyanins, and the mean degree of polymerization of tannins by HPLC by UV detection, total antioxidant capacity via spectrophotometric methods and chromatic characteristics via CIELAB.

OENOLOGICAL POTENTIAL OF AUTOCHTHONOUS SACCHAROMYCES CEREVISIAE STRAINS AND THEIR EFFECT ON THE PRODUCTION OF TYPICAL SAVATIANO WINES

Due to the global demand for terroir wines, the winemaking industry has focused attention on exploiting the local yeast microflora of each wine growing region to express the regional character and enhance the sensory profile of wines such as varietal typicity and aroma complexity. The objective of the present study was to isolate and compare the indigenous strains of Saccharomyces cerevisiae present in different vineyards in the Mesogeia – Attiki wine region (Greece), evaluate their impact on chemical composition and sensory profile of Savatiano wines and select the most suitable ones for winemaking process.