INVESTIGATION OF MALIC ACID METABOLIC PATHWAYS DURING ALCOHOLIC FERMENTATION USING GC-MS, LC-MS, AND NMR DERIVED 13C-LABELED DATA

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.

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.

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.

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).

In this work we briefly present a microfluidic device aiming to sort yeast cells according to their morphology. The technology is based upon microfluidic chips made out of Polydimethylsiloxane and glass using soft lithography processes and replica molding. The microfluidic device was used for encapsulating single yeast cells in liquid droplets containing growth medium. Liquid droplet containing yeast cells were sorted using a real time imaging and decision-making process.