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…

Grouping Vitis vinifera grapevine varieties based on their aromatic composition

Climate change is likely to impact wine typicity across the globe, raising concerns in wine regions historically renowned for the quality of their terroir1. Amongst several changes in viticultural practices, replacing some of the planting material (i.e. clones, rootstocks and cultivars) is thought to be one of the most promising potential levers to be used for adapting to climate change. But the change of cultivars also involves the issue of protecting the region’s wine typicity. In Bordeaux (France), extensive research has been conducted on identifying meridional varieties that could be good candidates to help guard against the effects of climate change2 while less research has been done concerning their impacts on Bordeaux wine typicity.

DEVELOPMENT OF BIOPROSPECTING TOOLS FOR OENOLOGICAL APPLICATIONS

Wine production is a complex biochemical process that involves a heterogeneous microbiota consisting of different microorganisms such as yeasts, bacteria, and filamentous fungi. Among these microorganisms, yeasts play a predominant role in the chemistry of wine, as they actively participate in alcoholic fermentation, a biochemical process that transforms the sugars in grapes into ethanol and carbon dioxide while producing additional by-products. The quality of the final product is greatly influenced by the microbiota present in the grape berry, and the demand for indigenous yeast starters adapted to specific grape must and reflecting the biodiversity of a particular region is increasing. This supports the concept that indigenous yeast strains can be associated with a “terroir”.

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.

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.

PHOTOCHEMICAL DEGRADATION OF TRYPTOPHAN IN MODEL WINE: IMPACT OF HEAVY METALS AND OXYGEN ON 2-AMINOACETOPHENONE FORMATION

The wine industry worldwide faces more and more challenges due to climate change, such as increased dryness in some areas, water stress, sunburn and early harvesting during hot summer temperatures¹. One of the resulting problems for the wine quality might be a higher prevalence of the untypical aging off-flavor (ATA)². A substance, which Rapp and Versini made responsible for ATA, is the 2-aminoace-tophenone (2-AAP)³. 2-AAP in wine causes a naphthalene, wet towels, wet wool, acacia flower or just a soapy note⁴.