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…

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.

Overhead spray water treatment as a mitigation strategy for reducing vine stress and preserving grape quality during heatwaves

Changes in climate have been influencing the quality of wine grapes worldwide. The impact of extreme climate events over short periods is increasingly recognized as a serious risk to grape quality and yield quantity. In this study the mitigation effects of a pulsed water spray on vine canopy during heatwave events has been evaluated for maintaining vine condition during the growing season and grape quality. Vines of three varieties (Malbec, Bonarda, and Syrah) under drip irrigation in the UNCuyo experimental vineyard were treated with an overhead pulsed water spray.

PREVALENCE OF OAK-RELATED AROMA COMPOUNDS IN PREMIUM WINES

Barrel fermentation and barrel-ageing of wine are commonly utilised practices in premium wine production. The wine aroma compounds related to barrel contact are varied and can enhance a range of wine aromas and flavours, such as ‘struck flint’, ‘caramel’, ‘red berry’, ‘toasty’ and ‘nutty’, as well as conventional oaky characters such as ‘vanilla’, ‘spice’, ‘smoky’ and ‘coconut’. A survey of commercially produced premium Shiraz, Cabernet Sauvignon, Pinot Noir and Chardonnay wines was conducted, assessing the prevalence of compounds that have been proposed as barrel-ageing markers¹ including oak lactones, volatile phenols, furanones, aldehydes, thiazoles2,3, phenylmethanethiol⁴ and 2-furylmethanethiol.⁵

UNTARGETED METABOLOMICS ANALYSES TO IDENTIFY A NEW SWEET COMPOUND RELEASED DURING POST-FERMENTATION MACERATION OF WINE

The gustatory balance of dry wines is centered on three flavors, sourness, bitterness and sweetness. Even if certain compounds were already identified as contributing to sweetness, some taste modifications remain largely unexplained1,2. Some empirical observations combined with sensory analyzes have shown that an increase of wine sweetness occurs during post-fermentation maceration³. This step is a key stage of red winemaking during which the juice is left in contact with the marc, that contains the solid parts of the grape (seeds, skins and sometimes stems). This work aimed to identify a new taste-active compound that contributes to this gain of sweetness.

FUNCTIONALIZED MESOPOROUS SILICA IS A VIABLE ALTERNATIVE TO BENTONITE FOR WINE PROTEIN STABILIZATION

The presence of grape-derived heat unstable proteins can lead to haze formation in white wines [1], an instability prevented by removing these proteins by adding bentonite, a hydrated aluminum silicate that interacts electrostatically with wine proteins leading to their flocculation. Despite effective, using bentonite has several drawbacks as the costs associated with its use, the potential negative effects on wine quality, and its environmental impact, so that alternative solutions are needed.