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…

STATISTICAL COMPARISON OF GROWTH PARAMETERS OF NINE BIOPROTECTION STRAINS IMPLEMENTED ON ARTIFICIALLY CONTAMINATED SYNTHETIC MUST

In recent years, consumer demand for products without chemical additives increased, becoming a priority for the wine sector. SO₂ is widely used for its multiple properties including antiseptics, antioxidants and antioxidasics and the strategy of bioprotection in winemaking represents now an alternative to this chemical additive. In oenology, results have highlighted the interest of bioprotection to limit the development of microorganisms like Hanseniaspora uvarum and thus reduce the doses of sulphite. Indeed, this species is considered because of its acetic acid and methyl butyl acetate production, the latter can cover the varietal character of wines.

WHICH IMPACT FOR PROANTHOCYANIDIC TANNINS ON RED WINE FRUITY AROMA? SENSORY AND PHYSICOCHEMICAL APPROACHES

Previous research on the fruity character of red wines highlighted the role of esters. Literature provides evidence that, besides these esters, other compounds that are not necessarily volatiles may have an important impact on the overall aroma of wine, contributing to a modulation of its global aromatic expression. The goal of this work was to assess the olfactory consequences of a mixture between esters and proanthocyanidic tannins, through sensory and physico-chemical approaches.
Sensory analysis of numerous aromatic reconstitutions, including triangular tests, detection thresholds, and sensory profiles, were conducted in order to evaluate the sensory impact of tannins on red wine esters perception.

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].

PRODUCTION OF A FUNCTIONAL BEVERAGE FROM WINEMAKING BY-PRODUCTS: A NEW WAY OF VALORISATION

In the challenge of transforming waste into useful products that can be re-used in a circular economy perspective, winery by-products can be considered as a source of potentially bioactive molecules such as polyphenols. The wine industry generates each year 20 million tons of by-products. Kombucha fermentation is an ancestral process which allow to increase the biological properties of tea by the action of a microbial consortium formed by yeasts and bacteria called SCOBY. It belongs to the field of healthy food for which the interest of consumers is growing. The objective of this work was to propose a new functional beverage made from winemaking by-products fermented by a Kombucha SCOBY.

HOW OXYGEN CONSUMPTION INFLUENCES RED WINES VOLTAMMETRIC PROFILE

Phenolic compounds play a central role in sensory characteristics of wine, such as colour, mouthfeel, flavour and determine its shelf life. Furthermore, the major non-enzymatic wine oxidation process is due to the catalytic oxidation of phenols in quinones. Due their importance, during the years have been developed different analytical methods to monitor the concentration of phenols in wine, such as Folin-Ciocalteu method, spectrophotometric techniques and HPLC. These methods can also be used to follow some oxidation-related chemical transformations.