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…

PROFILING OF LIPIDS IN WINES FROM MONOCULTURE FERMENTATION WITH INDIGENOUS METSCHNIKOWIA YEAST SPECIES

Lipids are a diverse group of organic compounds essential for living systems. They are vital compounds for yeast which makes them an important modulator of yeast metabolism in alcoholic fermentation. This study presents a comprehensive lipidome analysis of wine samples from the Vitis vinifera L., Maraština. The fermentation trails were set up in monoculture with different indigenous yeast strains selected from a collection of native yeasts established at the Institute for Adriatic Crops and Karst Reclamation in 2021, previously isolated from Croatian Maraština grapes: Metschnikowia pulcherrima, Metshnikowia sinensis/shanxiensis , and Metschnikowia chyrsoperlae.

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.

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.

POTENTIAL OF PEPTIDASES FOR AVOIDING PROTEIN HAZES IN MUST AND WINE

Haze formation in wine during transportation and storage is an important issue for winemakers, since turbid wines are unacceptable for sale. Such haze often results from aggregation of unstable grape proteinaceous colloids. To date, foreseeably unstable wines need to be treated with bentonite to remove these, while excessive quantities, which are often required, affect the wine volume and quality (Cosme et al. 2020). One solution to avoid these drawbacks might be the use of peptidases. Marangon et al. (2012) reported that Aspergillopepsins I and II were able to hydrolyse the respective haze-relevant proteins in combination with a flash pasteurisation. In 2021, the OIV approved this enzymatic treatment for wine stabilisation (OIV-OENO 541A and 541B).

SHIRAZ FLAVONOID EXTRACTABILITY IMPACTED BY HIGH AND EXTREME HIGH TEMPERATURES

Climate change is leading to an increase in average temperature and in the severity and occurrence of heatwaves, and is already disrupting grapevine phenology. In Australia, with the evolution of the weather of grape growing regions that are already warm and hot, berry composition including flavonoids, for which biosynthesis depends on bunch microclimate, are expected to be impacted [1]. These compounds, such as anthocyanins and tannins, contribute substantially to grape and wine quality. The goal of this research was to determine how flavonoid extraction is impacted when bunches are exposed to high (>35 °C) and extreme high (>45 °C) temperatures during berry development and maturity.