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…

Monitoring of Pesticide Residues from Vine to Wine

Those previous years, pesticides are often brought to the forefront by media. Questions arose about their toxicity for growers and consumers. Even if a downward trend is underway, the use of pesticides is required to ensure steady quality and quantity of harvests. A large number of active ingredients are authorized but regarding viticulture, mainly insecticides and fungicides are applied, to control pests and diseases and to increase crop yield. Some phytosanitary products, principally fungicides, applied close to the harvest date may frequently be detected in wines.

Impact of industrial-scale serial filtration on macromolecules in red wines

Filtration is a critical step in ensuring the clarity and microbial stability of wine prior to bottling. However the process of filtering potentially reduces red wine quality by removing some of the macromolecules that contribute to the texture of the wine. Commercial red wines, Cabernet Sauvignon (CAS) and Shiraz (SHZ), of two vintages and two grades (premium grade wines from the older vintage: CAS13 and SHZ13; and standard grade wines from a younger vintage: CAS14 and SHZ14) were filtered through industrial-scale commercial filtration units prior to bottling. Samples were taken before and after cross-flow filtration, lenticular filters, 0.65 µm and 0.45 µm pore size nylon membrane filters. The concentration and composition of macromolecules, including tannins and polysaccharides, were measured in all samples as well as particle size distribution and wine colour.

Impact of smoke exposure on the chemical composition of grapes

Vineyard exposure to smoke can lead to grapes and wine which exhibit objectionable smoky and ashy aromas and flavours, more commonly known as ‘smoke taint’ [1, 2]. In the last decade, significant bushfires have occurred around the world, including near wine regions in Australia, Canada, South Africa and the USA, as a consequence of the warmer, drier conditions associated with climate change. Considerable research has subsequently been undertaken to determine the chemical, sensory and physiological consequences of grapevine exposure to smoke. The sensory attributes associated with smoke-tainted wine have been linked to the presence of several smoke-derived volatile phenols, such as guaiacols, syringols and cresols [2].

Evaluation of colloidal stability in white and rosé wines investing Dynamic Light Scattering technology

Proteins constitute one of the three main components of grape juice and white wine, phenolic compounds and polysaccharides being the others. A specific group of the total grape-derived proteins resists degradation or adsorption during the winemaking process and remains in finished wine if not removed by the commonplace commercial practice of bentonite fining. While bentonite is effective in removing the problematic proteins, it is claimed to adversely affect the quality of the treated wine under certain conditions, through the removal of colour, flavor and texture compounds. A number of studies have indicated that different protein fractions require distinct bentonite concentrations for protein removal and consequent heat stabilization.

Reaction Mechanisms of Copper and Iron with Hydrogen Sulfide and Thiols in Model Wine

Fermentation derived sulfidic off-odors due to hydrogen sulfide (H2S) and low molecular weight thiols are commonly encountered in wine production and removed by Cu(II) fining. However, the mechanism underlying Cu(II) fining remains poorly understood, and generally results in increased Cu concentration that lead to deleterious reactions in finished wine. The present study describes a mechanistic investigation of the iron and copper mediated reaction of H2S, cysteine, 3-sulfanylhexan-1-ol, and 6-sulfanylhexan-1-ol with oxygen. The concentrations of H2S, thiols, oxygen, and acetaldehyde were monitored over time. It was found that Cu(II) was rapidly reduced by both H2S and thiols to Cu(I).