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…

FREE TERPENE RESPONSE OF ‘MOSCATO BIANCO’ VARIETY TO GRAPE COLD STORAGE

Temperature control is crucial in wine production, starting from grape harvest to the bottled wine storage. Climate change and global warming affect the timing of grape ripening, and harvesting is often done during hot summer days, influencing berry integrity, secondary metabolites potential, enzyme and oxidation phenomena, and even fermentation kinetics. To curb this phenomenon, pre-fermentative cold storage can help preserve the grapes and possibly increase the concentration of key secondary metabolites. In this study, the effect of grape pre-fermentative cold storage was assessed on the ‘Moscato bianco’ white grape cultivar, known for its varietal terpenes (65% of free terpenes represented by linalool and its derivatives) and widely used in Piedmont (Italy) to produce Asti DOCG wines.

AROMATIC AND FERMENTATIVE PERFORMANCES OF HANSENIASPORA VINEAE IN DIFFERENT SEQUENTIAL INOCULATION PROTOCOLS WITH SACCHAROMYCES CEREVISIAE FOR WHITE WINEMAKING

Hanseniaspora vineae (Hv) is a fermenting non-Saccharomyces yeast that compared to Saccharomyces cerevisiae (Sc) present some peculiar features on its metabolism that make it attractive for its use in wine production. Among them, it has been reported a faster yeast lysis and release of polysaccharides, as well as increased ß-glucosidase activity. Hv also produces distinctive aroma compounds, including elevated levels of fermentative compounds such as ß-phenylethyl acetate and norisoprenoids like safranal. However, it is known for its high nutritional requirements, resulting in prolonged and sluggish fermentations, even when complemented with Sc strain and nutrients.

WINE CONSUMER TRADE-OFF BETWEEN ORGANOLEPTIC CHARACTERISTICS AND SUSTAINABLE CLAIMS. AN EXPERIMENT ON RED WINES FROM BORDEAUX REGION

In economics, the perception of wine quality is not limited to sensorial characteristics: an indication of the region of production significantly affects the perception of quality and consumers’ WTP ([1]; [2]). However, [3] or more recently [4] show that even if a wine has an organic label, the taste of wine remains the predominant criterion in consumer preferences. The contribution of our experiment is to evaluate the impact of responsible attributes (organic label, Non Added Sulfites, HVE certification) on the appreciation of several red wines on the market. More than 280 consumers participated to the present study and they perform 25 tastings divided into 5 different sessions. 20 different red wines from Bordeaux Area are tasted.

RED WINE AGING WITHOUT SO₂: WHAT IMPACT ON MICROBIAL COMMUNITY?

Nowadays, the use of food preservatives is controversial, SO2 being no exception. Microbial communities have been particularly studied during the prefermentary and fermentation stages in a context of without added SO2. However, microbial risks associated with SO2 reduction or absence, particularly during the wine aging process, have so far been little studied. The microbiological control of wine aging is a key issue for winemakers wishing to produce wines without added SO2. The aim of the present study is to evaluate the impact of different wine aging strategies according to the addition or not of SO2 on the microbiological population levels and diversity.

PROTEOMIC STUDY OF THE USE OF MANNOPROTEINS BY OENOCOCCUS OENI TO IMPROVE MALOLACTIC FERMENTATION

Malolactic fermentation (MLF) is a desired process to decrease acidity in wine. This fermentation, carried out mostly by Oenococcus oeni, is sometimes challenging due to the wine stress factors affecting this lactic acid bacterium. Wine is a harsh environment for microbial survival due to the presence of ethanol and the low pH, and with limited nutrients that compromise O. oeni development. This may result in slow or stuck fermentations. After the alcoholic fermentation the nutrients that remain in the medium, mainly released by yeast, can be used in a beneficial way by O. oeni during MLF.