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…

THE EFFECT OF BENTONITE FINING ON THE VOLATILE AND NON-VOLATILE PROFILE OF ITALIAN WHITE WINES

Marselan wines have an unusual high proportion of seed derived tannins from grapes having high proportions of skins, which are rich in tannins. But the causes behind this characteristic have not yet been identified. In vintage 2023 wines were made at experimental scale (9 kg by experimental unit) from Arinarnoa, Marselan and Tannat Vitis vinifera grape cultivars by traditional maceration, and by techniques aimed to increase the wine content in skin derived tannin: addition of extraction enzymes, addition at vatting of grape-skin enological tannins, or by extended maceration, known to increase the seed derived tannin contents of wines. Macerations were of 7 days, except in the extended macerations that were of 15 days.

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).

IMPACT OF NEW BIO STIMULANTS ON GRAPE SECONDARY METABOLITES UNDER CLIMATE CHANGE CONDITIONS

In a context of climate change and excessive use of agrochemical products, sustainable approaches for environmental and human health such as the use of bio stimulants in viticulture represent a potential option, against abiotic and biotic threats. Bio stimulants are organic compounds, microbes, or a combination of both, that stimulate plant’s vital processes, allowing high yields and good quality products. In vines, may trigger an innate immune response leading to the synthesis of secondary metabolites, key compounds for the organoleptic properties of grapes and wines.

NEW INSIGHTS INTO THE EFFECT OF TORULASPORA DELBRUECKII/SACCHAROMYCES CEREVISIAE INOCULATION STRATEGY ON MALOLACTIC FERMENTATION PERFORMANCE

Winemaking is influenced by micro-organisms, which are largely responsible for the quality of the product. In this context, Non-Saccharomyces and Saccharomyces species are of great importance not only because it influences the development of alcoholic fermentation (AF) but also on the achievement of malolactic fermentation (MLF). Among these yeasts, Torulaspora delbrueckii allows in sequential inoculation with strains of S. cerevisiae shorter MLF realizations [5] . Little information is available on the temporal effect of the presence of T. delbrueckii on (i) the evolution of AF and (ii) the MLF performance.

CHARACTERISTIC EXTRACTION OF THE PHENOL COMPOUNDS IN KOSHU (VITIS VINIFERA CV.) WINE DURING THE MACERATION

Koshu is one of the indigenous grape variety that has been grown in Japan for more than one thousand years. Recent research showed that it has 70% of Vitis vinifera genes. In 2010, the Koshu variety was included in ‘International List of Vine and Varieties and their Synonyms’ managed by the ‘International Organisation of Vine and Wine’ and has further fueled its popularity in Japan. It is the most cultivated variety for winemaking in Japan.
Koshu berries have light purple skins. The variety is mainly used to produce white wines such as an aromatic wine and a wine produced by sur lie method although various styles are produced.