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…

ANTHOCYANINS EXTRACTION FROM GRAPE POMACE USING EUTECTIC SOLVENTS

Grape pomace is one of the main by-products generated after pressing in winemaking.Emerging methods, such as ultrasound-assisted extraction with eutectic mixtures, have great potential due to their low toxicity, and high biodegradability. Choline chloride (ChCl) was used as a hydrogen bond acceptor and its corresponding hydrogen bond donor (malic acid, citric acid, and glycerol: urea). Components were heated at 80 °C and stirred until a clear liquid was obtained. Distilled water was added (30 % v/v). A solid-liquid ratio of 1 g pomace per 10 ml of eutectic solvent was used.

OENOLOGICAL STRATEGIES FOR THE REMOVAL OF PINKING IN WHITE WINE

The pinking of in white wine is the turning of color from yellow to salmon hue. White wines obtained from certain grape varieties (e.g. Chardonnay, Sauvignon blanc, Riesling, Trebbiano di Lugana) showed to be susceptible to pinking [1] that has been evaluated by an assay providing the addition of hydrogen peroxide. Even if its appearance does not seem to affect the sensory properties [2], strategies are necessary for its removal. Nowadays, the treatment with polyvinylpolipirroline (PVPP) was reported to significantly decrease the pink color [3].

WINE FERMENTATION METABOLITES PRODUCED BY TWO TORULASPORA DELBRUECKII STRAINS ISOLATED FROM OKANAGAN VALLEY, BC, CANADA VINEYARDS

Wine aroma is influenced by various factors, from agricultural practices in the vineyard to the enological choices made by winemakers throughout the vinification process. Spontaneous fermentations have a characteristically deeper complexity of aromas when compared to fermentations that have been inoculated with Saccharomyces (S.) cerevisiae because of the diversity of microflora naturally present on grape skins. Non-Saccharomyces yeast are being extensively studied for their ability to positively contribute to wine aroma and flavour. These yeasts are known to liberate more bound volatile compounds present in grape must than S. cerevisiae through the enzymatic action of β-glucosidases and β-lyases1.

WINE SWIRLING: A FIRST STEP TOWARDS THE UNLOCKING OF THE WINE’STASTER GESTURE

Right after the pouring of wine in a glass, a myriad of volatile organic compounds, including ethanol, overwhelm the glass headspace, thus causing the so-called wine’s bouquet [1]. Otherwise, it is worth noting that during wine tasting, most people automatically swirl their glass to enhance the release of aromas in the glass headspace [1]. About a decade ago, Swiss researchers revealed the complex fluid mechanics underlying wine swirling [2]. However, despite mechanically repeated throughout wine tasting, the consequences of glass swirling on the chemical space found in the headspace of wine glasses are still barely known.

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.