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…

Effect of supplementation with inactive yeast during alcoholic fermentation in base wine for sparkling

INTRODUCTION: Foam stability of sparkling wines is significantly favored by the presence of surface active agents such as proteins and polysaccharides [1]. For that reason, the renowned sparkling wines are aged after the second fermentation in contact with the lees for several months (even years). Thereby wines are enriched in these macromolecules due to yeast autolysis. Since this practice is slow and costly, winemakers are seeking for alternative procedures to increase their concentration in base wines. In that sense, the supplementation with inactive yeast during alcoholic fermentation has been proposed [2]. The aim of this study was to determine whether this new strategy is really useful for enriching base wines in macromolecules and for improving foam properties of the base wines.

Wood from barrique: release of phenolic compounds and permeability to oxygen

Chemical and sensory changes occurring in red wine during ageing in oak barrique are due to the slow and gradual entrance of oxygen along with a release of ellagic tannin from the wood. Though oxygen can enter the cask through the bunghole, it is not clear the role of permeation through the wood staves as well as the amount of oxygen entering by permeation. The distribution of the released ellagic tannins in the wine ageing is also unknown. The oxygen passing through the bunghole may have a different wine ageing effect compared to the oxygen permeating through the wooden staves owing to the uneven ellagic tannin concentration throughout the wine.

Quantification of the production of hydrogen peroxide H2O2 during wine oxidation

Chemical studies aiming at assessing how a wine reacts towards oxidation usually focus on the characterization of wine constituents, such as polyphenols, or oxidation products. As an alternative, the key oxidation intermediate hydrogen peroxide H2O2 has never been quantified, although it plays a pivotal role in wine oxidation. H2O2 is obtained from molecular oxygen as the result of a first cascade of oxidation reactions involving metal ions and polyphenols. The produced H2O2 then reacts in a second cascade of oxidation to produce reactive hydroxyl radicals that can attack almost any chemical substrate in wine.

Effect of nanofiltration on the chemical composition and wine quality

In Enology the conventional processes of filtration for clarification and stabilization are giving place to alternative membrane processes, including nanofiltration (NF). Furthermore, the increased alcohol content in wines recorded in recent years became an important issue for all the main wine producing countries. Among techniques available to the wine industry to reduce the ethanol content, NF is certainly one of the newest. This study is focused on the evaluation of NF influence on wine physical-chemical composition, including mineral content, which in accordance to our best knowledge is a novelty.

Ripening of cv. Cabernet Sauvignon grapes: polysaccharides fractions evolution and phenolic extractability

Polysaccharides and more specifically pectins, make up a significant portion of the cell wall material of the plant cells including the grapes. During the fruit ripening the associated softening is related to the breakdown of the cell wall polysaccharides. During this process, it is expected that polysaccharides that are soluble in red wine will be formed influencing its texture. Anthocyanins are responsible for the wine color and tannins for the astringency, body and bitterness of the wine. In the skins, these compounds are located in the cell vacuoles and the barrier that conditions their extractability is the skin cell wall that may determine the mechanical resistance, the texture and the ease of processing berries. The aim of this work was study the evolution of the polysaccharides and the anthocyanin and tannin extractability during the ripening period in Cabernet Sauvignon grapes, trying to correlate these variables.