Macrowine 2021
IVES 9 IVES Conference Series 9 Directed Evolution of Oenococcus oeni: optimising yeast-bacteria interactions for improved malolactic fermentation

Directed Evolution of Oenococcus oeni: optimising yeast-bacteria interactions for improved malolactic fermentation

Abstract

Malolactic fermentation (MLF) is a secondary step in the vinification process and it follows alcoholic fermentation (AF) which is predominantly carried out by Saccharomyces cerevisiae. These two processes result in the degradation of metabolites to produce secondary metabolites which also contribute to the final wine flavour and quality. AF results in the production of ethanol and carbon dioxide from sugars and MLF stems from the degradation of L-malic acid (a dicarboxylic acid) to L-lactic acid (a monocarboxylic acid). The latter process results in a smoother texture as the acidity of the wine is reduced by the process, it also adds to the flavour complexity of the wine. The species responsible for this fermentation step belong to the Pediococci, Lactobacilli and Oenococci genera. Only Oenococcus oeni and Lactobacillus plantarum have been commercialised. The former is the dominant species that is often found in both spontaneous and inoculated fermentations. In spite of inoculation MLF is quite unstable and a successful fermentation is not always guaranteed. Sluggish or stuck fermentations may occur due to many physico-chemical factors. Also, the interactions between the yeast and bacteria during the vinification process play an important role in the success of MLF. Therefore, appropriate selection of strains is important, unfortunately selecting strains is time consuming and limited only to specific winemaking conditions. To overcome this, research has investigated strain improvement, however recombinant technology is controversial. The use of non-recombinant techniques such as mutagenesis, hybridisation and Directed Evolution has become popular. The aim of this study is to optimise yeast-bacteria interactions by use of Directed Evolution as a tool to improve lactic acid bacteria, in this way, try and guarantee the success of MLF. Two S. cerevisiae strains (Cross Evolution® and EC1118®) were used as selective pressures for O oeni S5 populations. The bacterial populations were exposed to synthetic wine fermentations for 30 and 50 generations after which 30 bacterial isolates were evolved using both yeast and were characterised for fermentation efficacy. The results show that the general performance of the isolates was improved in comparison to the parental strain. Only 3 isolates after 30 generations showed a specific improvement when inoculated with ‘driver’ yeast than with other yeast strains. After 50 generations all the strains showed improvement in terms of fermentation rates, but not all strains had a higher fermentation efficacy in comparison to the parent strain. This study shows the potential of Directed Evolution as a tool for strain improvement using a biotic selective pressure as opposed to physico-chemical selective pressures. It also, shows the possibility of improving yeast-bacteria interactions by having a tailor-made pair for successful AF and MLF.

Publication date: May 17, 2024

Issue: Macrowine 2016

Type: Poster

Authors

Seipati Tenyane*, Debra Rossouw, Florian Bauer

*Stellenbosch Universiy

Contact the author

Tags

IVES Conference Series | Macrowine | Macrowine 2016

Citation

Related articles…

Metabolomics comparison of non-Saccharomyces yeasts in Sauvignon blanc and Shiraz

Saccharomyces cerevisiae (SC) is the main driver of alcoholic fermentation however, in wine, non-Saccharomyces species can have a powerful effect on aroma and flavor formation. This study aimed to compare untargeted volatile compound profiles from SPME-GC×GC-TOF-MS of Sauvignon blanc and Shiraz wine inoculated with six different non-Saccharomyces yeasts followed by SC. Torulaspora delbrueckii (TD), Lachancea thermotolerans (LT), Pichia kluyveri (PK) and Metschnikowia pulcherrima (MP) were commercial starter strains, while Candida zemplinina (CZ) and Kazachstania aerobia (KA), were isolated from wine grape environments. Each fermentation produced a distinct chemical profile that was unique for both grape musts. The SC-monoculture and CZ-SC sequential fermentations were the most distinctly different in the Sauvignon blanc while the LT-SC sequential fermentations were the most different from the control in the Shiraz fermentations.

Novel analytical technologies for wine fingerprinting in and beyond the laboratory

For characterization, sensory designing and authentication rapid analytical technologies have become available. Some, like Proton Transfer Reaction Mass Spectrometry allow a rapid spectrum of the volatile compounds of wines. Combined with chemometrics wines can be characterized. The same approach can be used to calculate the results of virtual mixtures and allow formulation of constant quality blends. Other new techniques and portable devices based on spectroscopy allow measurements on production sites and in grocery stores, even for the smart consumer. We will present some examples of the application of these techniques for authentication of wines, both in the laboratory and on site.

Testing the effectiveness of Cell-Wall material from grape pomace as fining agent for red wines

Lately several works highlighted the capacity of grape cell-wall material (CWM) to interact with proanthocyanidins (PA), indicating its potential use as fining agent for red wines.1–4 However, those studies were performed by using purified PAs and very high doses of CWM (almost ten-fold higher than those used in wine industry for other commercial fining agents). The present study focuses on the applicability of CWM from Cabernet sauvignon pomace as fining agent for red wines under real winery conditions. Grapes of cultivar Cabernet sauvignon were harvested at three different maturity levels
(unripe, mature, and overripe) and used for red winemaking. The pomace of such vinifications were used as source of CWM, and applied into red wines at two different concentrations: 0.2 g/L and 2.5 g/L.

Effects of a new vacuum evaporation method on chemical and sensory properties of must and wine

A new process for vacuum evaporation was developed where evaporation takes place near the inner surface of a vortex produced by a rotor submerged in the liquid. Contrary to the state of the art the Vortex rotor process does not need a vacuum vessel but the rotating liquid creates a geometrically stable low pressure void surrounded by a vortex stabilized by the equilibrium between centrifugal forces and the pressure difference. First tests with water and sugar solutions at concentrations similar to grape must were conducted to verify the theoretical predictions, test the performance under different conditions and study the effect of various process parameters (Rösti et al 2015).

Defining the mechanisms and impact of winemaking treatments on tannin and polysaccharides in red wine: recent progress in creating diverse styles

Tannin and polysaccharide concentration and composition is important in defining the texture of red wines, but can vary due to factors such as cultivar, region, grape ripeness, viticultural practices and winemaking techniques. However, the concentration and composition of these macromolecules is dependent not only on grape tannin and polysaccharide concentration and composition, but also their extractability and, in the case of polysaccharides, their formation by yeast. Through studies into the influence of grape maturity, winemaking and sensory impacts of red grape polysaccharides, seed and skin tannins, recent research in our laboratory has shown that the processes involved in the extraction of these macromolecules from grapes and their retention in wine are very complex.