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 mixed Torulaspora delbrueckii-Saccharomyces cerevisiae culture on rose quality wine

Alcoholic fermentation using no Saccharomyces wine is an effective means of modulating wine aroma. This study investigated the impact of coinoculating Torulaspora delbruecki with two Saccharomyces cerevisiae commercial yeast (QA23, Lallemand; Red Fruit, Sepsa-Enartis) on enological quality parameters, volatile composition and sensory analysis. The following assays were performed on Tempranillo variety: Saccharomyces QA23 (CTQA), Saccharomyces Red Fruit (CTRF), coinoculated T. delbrueckii + S.cerevisiae QA23 (CIQA) and coinoculated T. delbrueckii + S.cerevisiae (CIRF).

Comprehensive exploration of wine aroma-related compounds as promoted by alternative vinification procedures in case of Zelen (Vitis vinifera L.) grapes processing

Not only vintner’s decisions in the vineyard, but also winemaker’s choices of technology approaches in the cellar play a significant role in the final wine style and quality. Whereas traditional technologies within chosen terroir are quite well explored and thus somehow predictable, there is no proper knowledge available on possible outcomes in case of implementing novel, alternative winemaking strategies. To reveal their effects on wine aroma compounds and sensory characteristics, two alternative strategies
(cryoextraction or addition of whole grape berries during last stages of fermentation) were compared to classical Vipava valley winemaking approach as normally used for an autochthonous variety Zelen. After separate vinification and bottling, all the experimental wines were subjected to semiquantitative metabolic profiling of volatile compounds (VOCs) by means of GC/MS and were then also sensorialy evaluated by pre-trained panel.

Correlations between sensory characteristics and colloidal content in dry white wines

Must clarification is an important step occurring just after grape extraction in the elaboration of white wine, consisting in a solid-liquid separation. Traditionally, low must turbidity, around 50-150 NTU, is generally reached in white winemaking in order to prevent reductive aromas and facilitating alcoholic fermentation. Alternatively, a higher turbidity (300 NTU or above) can be sought for reasons such as a better expression of grapes identity (terroir), or for getting a must matrix that could supposedly lead to wines having greater ageing potential.

Study of the content of amino acids and biogenic amines in sparkling red wines

The production of red sparkling wines is lower in Spain in comparison with the winemaking of white or rosé sparkling wines. In red sparkling wine processing it is essential to obtain suitable base wines that should have moderate alcohol content, high acidity, good color values, an adequate mouth-feel and a sweet tannin. Grapes for sparkling wine production have to be harvested at low maturity stages, with lower alcohol contents and higher acidities, which will that the phenolic maturity of the grapes is also low, showing green tannins. This paper analyses different treatments in order to minimize these inconveniences: cold maceration-prefermentation and delestage to elaborate the grapes with lower maturity, must nanofiltration, and the partial osmosis of the wines made from grapes with an adequate maturity degree.

Metabolomic profile of red non-V. vinifera genotypes

Vitis vinifera L. is the most widely cultivated Vitis species which includes numerous cultivars. Owing to their superior quality of grapes, these cultivars were long considered the only suitable for the production of fine wines. However, the lack of resistance genes in V. vinifera against major grapevine pathogens, requires for its cultivation frequent spraying with large amount of fungicides. Thus, the search for alternative and more sustainable methods to control the grapevine pathogens have brought the breeders to focus their attention on other Vitis species. In fact, wild Vitis genotypes present multiple resistance traits against pathogens, such as powdery mildew, downy mildew and phylloxera.