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…

INTENSE PULSED LIGHT FOR VINEYARD WASTEWATER: A PROMISING NEW PROCESS OF DEGRADATION FOR PESTICIDES

The use of pesticides for vine growing is responsible for generating an important volume of wastewater. In 2009, 13 processes were authorized for wastewater treatment but they are expensive and the toxicological impact of the secondary metabolites that are formed is not clearly established. Recently photodecomposition processes have been studied and proved an effectiveness to degrade pesticides and to modify their structures (Maheswari et al., 2010, Lassale et al., 2014). In this field, Pulsed Light (PL) seems to be an interesting and efficient process (Baranda et al., 2017). Therefore, the aim of this work was to investigate the PL technology as a new process for the degradation of pesticides.

‘TROPICAL’ POLYFUNCTIONAL THIOLS AND THEIR ROLE IN AUSTRALIAN RED WINES

Following anecdotal evidence of unwanted ‘tropical’ character in red wines resulting from vineyard interventions and a subsequent yeast trial observing higher ‘red fruit’ character correlated with higher thiol concentrations, the role of polyfunctional thiols in commercial Australian red wines was investigated.
First, trials into the known tropical thiol modulation technique of foliar applications of sulfur and urea were conducted in parallel on Chardonnay and Shiraz.1 The Chardonnay wines showed expected results with elevated concentrations of 3-sulfanylhexanol (3-SH) and 3-sulfanylhexyl acetate (3-SHA), whereas the Shiraz wines lacked 3-SHA. Furthermore, the Shiraz wines were described as ‘drain’ (known as ‘reductive’ aroma character) during sensory evaluation although they did not contain thiols traditionally associated with ‘reductive’ thiols (H2S, methanethiol etc.).

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.

PAIRING WINE AND STOPPER: AN OLD ISSUE WITH NEW ACHIEVEMENTS

The sensory characteristics of wine are a topic studied by several researchers over time, but it continues to be a current and challenging subject. These characteristics are fundamental for the consumer acceptability, which has increasingly aroused their interest to modulate them in line with current market trends and innovation demands. The wine physical-chemical and sensory properties depend on a wide set of factors: they begin to be designed in the vineyard and are later constructed during the various stages of winemaking. Afterwards, the wine is placed in bottles and stored or commercialized.

CONSUMER PERCEPTION OF INTERSPECIFIC HYBRID RED WINE COLOR IN RELATION TO ANTHOCYANIN PROFILE AND CHEMICAL COLOR PARAMETERS

Interspecific hybrid winegrapes are of growing interest in the context of climate change based on their disease resistance and cold hardiness. In addition to a need for increased understanding of their chemical composition, there is little empirical evidence on the consumer perception of non-vinifera wine. Phenolic compounds, and particularly color, play an important organoleptic and quality determination role in wine, but can vary significantly in interspecific hybrid wines compared to wines produced from Vitis vinifera cultivars [1, 2, 3]. Anecdotally, the variation in anthocyanin species, interactions, and concentrations in interspecific hybrids could result in a variance from“vinifera-like” wine color.