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…

Non-invasive headspace sorptive extraction for monitoring volatile compounds production by saccharomyces and non-saccharomyces strains throughout alcoholic fermentation

Wine is a solution containing abundant volatile compounds which contribute to their aroma. Many of them are produced by yeast as metabolism by-products. Different yeast strains produce different volatile profiles. The possibility of studying the evolution of volatile compounds during fermentation, using sampling methods that not alter the volume of fermentation media, is of great interest. In spite of this, non-invasive methods to monitoring the evolution of volatile profile during fermentation have been seldom used. The goals of this work were to use by first time the headspace sorptive extraction (HSSE) as non-invasive method to monitor the evolution of volatile profiles throughout alcoholic fermentation and to study the changes on volatile profiles produced by Saccharomyces cerevisiae and Lachancea thermotolerans during fermentation of a must with high sugar content.

The impact of branched chain and aromatic amino acids on fermentation kinetics and aroma biosynthesis by wine yeast Saccharomyces cerevisiae

One of the major determinants of wine quality is the aroma. Wine aroma is the human perception of the matrix of grape and yeast derived volatiles and their interaction that contribute to flavour wine. Most common are higher alcohols, ester and aldehydes. In previous studies the formation of characteristic volatile compounds have been linked to the metabolism of branched-chain and aromatic amino acids
(BCAAs) in synthetic grape must. Here we report on an investigation to assess the impact of the initial amino acid concentration on the production of aroma compounds by the industrial yeast VIN13 grown in both synthetic and real grape musts.

How pressing techniques affect must composition and wine quality of Pinot blanc

This study investigates how the sensory profile of Pinot Blanc is affected from different maceration and pressing techniques. Grapes were sourced from four vineyards in the village Tramin in South Tyrol. For the experiment 200 kg of grapes from each vineyard site were hand picked the day before harvest for the commercial winery took place. Grapes were stored over night at 4°C, homogenized and processed in the experimental winery at Laimburg research centre the day after harvest. Four different pressing techniques were applied in duplicates of 100kg each.

Improving the phenolic composition of cv tempranillo wines by blending grapes of different ripening state

The aim of this work was to reduce the alcohol content of Tempranillo wine. Tempranillo wines were produced by grapes harvested at different ripening dates (August 11 which was 21 oBrix and September 28 with 25 oBrix). At the second date, the Tempranillo wines were elaborated as follows: grapes were destemmed, crushed and collected into 50 L stainless-steel vats. Before preferementative maceration in cold, 50 % (M1) and 70 % (M2) of the must have been replaced by the same percentage of must from the first harvest. In addition, a control wine (C) was performed with only grapes from the second harvest.

The effect of Nitrogen and Sulphur foliar applications in hot climates

ine nitrogen deficiency can negatively influence the aroma profile and ageing potential of white wines. Canopy management can alter vine microclimate, affect the nitrogen availability and influence the response of leaf senescence. Increasing the nitrogen availability to vines can increase the Yeast Assimilable Nitrogen (YAN) levels in harvested fruit and wine. Studies show that foliar nitrogen and sulphur applications at véraison, on low YAN Sauvignon blanc grapes have an effect on the level of amino acids (Jreij et al. 2009) and on S-containing compounds such as glutathione and thiols (Lacroux et al. 2008), which in turn can influence the formation of major volatiles and the aroma profile of the wine.