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…

IDENTIFYING POTENTIAL CHEMICAL MARKERS RESPONSIBLE FOR THE PERMISSIVENESS OF BORDEAUX RED WINES AGAINST BRETTANOMYCES BRUXELLENSIS USING UNTARGETED METABOLOMICS

All along the red winemaking process, many microorganisms develop in wine, some being beneficial and essential, others being feared spoilers. One of the most feared microbial enemy of wine all around the world is Brettanomyces bruxellensis. Indeed, in red wines, this yeast produces volatile phenols, molecules associated with a flavor described as “horse sweat”, “burnt plastic” or “leather”. To produce significant and detectable concentrations of these undesired molecules, the yeasts should first grow and become numerous enough. Even if the genetic group of the strain present and the cellar temperature may modulate the yeast growth rate¹ and thus the risk of spoilage, the main factor seems to be the wines themselves, some being much more permissive to B. bruxellensis development than others.

LARGE-SCALE PHENOTYPIC SCREENING OF THE SPOILAGE YEAST BRETTANOMYCES BRUXELLENSIS: UNTANGLING PATTERNS OF ADAPTATION AND SELECTION, AND CONSEQUENCES FOR INNOVATIVE WINE TREATMENTS

Brettanomyces bruxellensis is considered as the main spoilage yeast in oenology. Its presence in red wine leads to off-flavour due to the production of volatile phenols such as 4-vinylphenol, 4-vinylguaiacol, 4-ethylphenol and 4-ethylguaiacol, whose aromatic notes are unpleasant (e.g. animal, leather, horse or pharmaceutical). Beside wine, B. bruxellensis is commonly isolated from beer, kombucha and bioethanol production, where its role can be described as negative or positive. Recent genomic studies unveiled the existence of various populations.

HOW DOES ULTRASOUND TREATMENT AFFECT THE AGEING PROFILE OF AN ITALIAN RED WINE?

Many wine styles require moderate or extended ageing to ensure optimal consumer experience. However, few consumers have the interest or ability to age wine themselves, and holding wine in optimal conditions for extended periods is expensive for producers. A study was conducted on the use of ul-trasound energy on wine, with particular reference to its impact on sensory and chemical profiles. The OIV has authorised the use of ultrasound for processing crushed grapes (must) in Resolution OENO 616-2019, but not yet for finished wine1,2.

CHARACTERIZATION OF ENOLOGICAL OAK TANNIN EXTRACTS BY MULTI-ANALYTICAL METHODS APPROACH

Oak tannin extracts are commonly used to improve wine properties. The main polyphenols found in oak wood extracts are ellagitannins¹ that release ellagic acid upon hydrolysis and comprise numerous structures². Moreover, oak tannin extracts contain other compounds giving a complex mixture. Consequently, the official OIV method based on gravimetric analysis of the tannin fraction adsorbed on polyvinylpolypyrrolidone is not sufficient to describe their composition and highlight their chemical diversity.

Managing changes in taste: lessons from champagne in britain 1800-1914

This paper focuses on how taste in wine (and other foods) changes and the implications of this process
for producers and merchants.
It draws primarily on the changing taste of and taste for champagne in Britain in the 19th century. Between 1850 and 1880 champagne went from a dosage level of around 20% (20 grams sugar / litre) to 0%. Champagne became the ‘dinner wine of the elite – drunk with roast meat and savoury dishes.
Contemporaries accepted that while most people could distinguish the taste of good champagne from that of bad, very few could distinguish very good from good.