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…

WHAT’S FUTURE FOR SANTORINI’S VITICULTURE IN THE CONTEXT OF CLIMATE CHANGE

The own-rooted vineyard of Santorini is a unique case of vineyard worldwide that is been cultivated for thousands of years. On the island’s volcanic soil, the vines are still cultivated with traditional techniques, which are adapted to the specific and extreme weather conditions that prevail on it. While climate change is a reality in the Mediterranean region, will Santorini vineyard endure its impact? The study of the traditional training systems, techniques and vine density, as well as the application of sustainable solutions (cover crops and use of kaolin etc.) revealed sustainable methods for the adaptation of the local viticulture to new climatic phenomena that tend to be more and more frequent in the region due to climate change.

ACIDIC AND DEMALIC SACCHAROMYCES CEREVISIAE STRAINS FOR MANAGING PROBLEMS OF ACIDITY DURING THE ALCOHOLIC FERMENTATION

In a recent study several genes controlling the acidification properties of the wine yeast Saccharomyces cerevisiae have been identified by a QTL approach [1]. Many of these genes showed allelic variations that affect the metabolism of malic acid and the pH homeostasis during the alcoholic fermentation. Such alleles have been used for driving genetic selection of new S. cerevisiae starters that may conversely acidify or deacidify the wine by producing or consuming large amount of malic acid [2]. This particular feature drastically modulates the final pH of wine with difference of 0.5 units between the two groups.

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”.

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).

EFFECTS OF LEAF REMOVAL AT DIFFERENT BUNCHES PHENOLOGICAL STAGES ON FREE AND GLYCOCONJUGATE AROMAS OF SKINS AND PULPS OF TWO ITALIAN RED GRAPES

Canopy-management practices are applied in viticulture to improve berries composition and quality, having a great impact on primary and secondary grape metabolism. Among these techniques, cluster zone leaf removal (defoliation) is widely used to manage air circulation, temperature and light radiation of grape bunches and close environment. Since volatiles are quantitatively and qualitatively influenced by the degree of fruit ripeness, the level of solar exposure, and the thermal environment in which grapes ripen, leaf removal has been shown to affect volatile composition of grape berries [1].