Project description:CGH analysis of copy number variation in 11 strains of Oenococcus oeni compared to strain AWRIB515. strain vs AWRIB515. One replicate per strain.
Project description:Pangenomic oligonucleotide expression array designed for exploring the transcriptome of O. oeni strains cultured in standard and wine stress conditions.
Project description:The correct development of malolactic fermentation depends on the capacity of Oenococcus oeni to survive under harsh wine conditions. In this study it was characterized the transcriptomic response of O. oeni PSU-1 during the adaptation to wine-like medium (WLM). The period evaluated was from the inoculation into WLM (12% v/v pH 3.4) until the beginning of malolactic fermentation (8h after inoculation).
Project description:Oenococcus oeni is the main lactic acid bacterium that carries out the malolactic fermentation in virtually all red wines and in some white and sparkling wines. O. oeni possesses an array of metabolic activities which can modify the taste and aromatic properties of wine. There is therefore industrial interest in the proteins involved in these metabolic pathways and related transport systems of this bacterium. In this work, we report the characterization of the O. oeni ATCC BAA-1163 proteome. Total and membrane protein preparations from O. oeni were standardized and analyzed by two-dimensional gel electrophoresis. Using tandem mass spectrometry we identified 226 different polypeptides corresponding to 155 unique proteins, which have been classified by their putative function and subjected to bioinformatics analysis.
Project description:Malolactic fermentation (MLF) positively influences the quality of the wine, and it occurs as a result of a lactic acid bacteria’s metabolism, mainly of the Oenococcus oeni species. However, delays and halting of MLF are frequent problems in the wine industry. This is mainly because O. oeni’s development is inhibited by different kinds of stress. Even though the sequencing of the genome of the PSU-1 strain of O. oeni, as well as other strains, has made it possible to identify genes involved in the resistance to some types of stress, all of the factors that could be involved are still unknown. With the aim of contributing to this knowledge, the random mutagenesis technique was used in this study as a strategy for genetic improvement of strains of the O. oeni species. The technique proved to be capable of generating a different and improved strain when compared to the PSU1 strain (the parent from which it descends). Then, we evaluated the metabolic behavior of both strains in three different wines. We used synthetic MaxOeno wine (pH 3.5; 15% v/v ethanol), red wine (Cabernet Sauvignon) and white wine (Chardonnay). Furthermore, we compared the transcriptome of both strains, grown in MaxOeno synthetic wine. The specific growth rate of the E1 strain was on average 39% higher in comparison to the PSU1 strain. Interestingly, E1 strain showed an overexpression of the OEOE_1794 gene, which encodes a UspA-like protein, which has been described as promoting growth. We observed that the E1 strain was able to convert, on average, 34% more malic acid into lactate than the PSU1 strain, regardless of the wine being used. On the other hand, the E1 strain showed a flux rate of fructose-6-phosphate production that was 86% higher than the mannitol production rate, and the internal flux rates increase in the direction of pyruvate production. This coincides with the higher number of OEOE_1708 gene transcripts observed in the E1 strain grown in MaxOeno. This gene encodes for an enzyme fructokinase (EC 2.7.1.4) involved in the transformation of fructose to fructose-6-phosphate. It is necessary to continue studying these genes in order to verify their biological function in O. oeni and their impact on MLF.