Microbial co-culturing strategies for fructo-oligosaccharide production.

Fructo-oligosaccharide (FOS) mixtures produced by fermentation contain large amounts of non-prebiotic sugars. Here we propose a mixed culture of Aureobasidium pullulans and Saccharomyces cerevisiae cells to produce FOS and consume the small saccharides simultaneously, thereby increasing FOS purity in the mixture. The use of immobilised A. pullulans in co-culture with encapsulated S. cerevisiae, inoculated after 10 h fermentation, enhanced FOS production in a 5 L bioreactor. Using this strategy, a maximal FOS concentration of 119 g L-1, and yield of 0.59 gFOS gsucrose-1, were obtained after 20 h fermentation, increasing FOS productivity from about 4.9 to 5.9 gFOS L-1 h-1 compared to a control fermentation of immobilized A. pullulans in monoculture. In addition, the encapsulated S. cerevisiae cells were able to decrease the glucose in the medium to about 7.6% (w/w) after 63 h fermentation. This provided a final fermentation mixture with 2.0% (w/w) sucrose and a FOS purity of over 67.0% (w/w). Moreover, a concentration of up to 58.0 g L-1 of ethanol was obtained through the enzymatic transformation of glucose. The resulting pre-purified FOS mixture could improve the separation and purification of FOS in downstream treatments, such as simulated moving bed chromatography.

New integrative computational approaches unveil the Saccharomyces cerevisiae pheno-metabolomic fermentative profile and allow strain selection for winemaking.

During must fermentation by Saccharomyces cerevisiae strains thousands of volatile aroma compounds are formed. The objective of the present work was to adapt computational approaches to analyze pheno-metabolomic diversity of a S. cerevisiae strain collection with different origins. Phenotypic and genetic characterization together with individual must fermentations were performed, and metabolites relevant to aromatic profiles were determined. Experimental results were projected onto a common coordinates system, revealing 17 statistical-relevant multi-dimensional modules, combining sets of most-correlated features of noteworthy biological importance. The present method allowed, as a breakthrough, to combine genetic, phenotypic and metabolomic data, which has not been possible so far due to difficulties in comparing different types of data. Therefore, the proposed computational approach revealed as successful to shed light into the holistic characterization of S. cerevisiae pheno-metabolome in must fermentative conditions. This will allow the identification of combined relevant features with application in selection of good winemaking strains.

Application of a high-throughput process analytical technology metabolomics pipeline to Port wine forced ageing process.

Metabolomics aims at gathering the maximum amount of metabolic information for a total interpretation of biological systems. A process analytical technology pipeline, combining gas chromatography-mass spectrometry data preprocessing with multivariate analysis, was applied to a Port wine "forced ageing" process under different oxygen saturation regimes at 60°C. It was found that extreme "forced ageing" conditions promote the occurrence of undesirable chemical reactions by production of dioxane and dioxolane isomers, furfural and 5-hydroxymethylfurfural, which affect the quality of the final product through the degradation of the wine aromatic profile, colour and taste. Also, were found high kinetical correlations between these key metabolites with benzaldehyde, sotolon, and many other metabolites that contribute for the final aromatic profile of the Port wine. The use of the kinetical correlations in time-dependent processes as wine ageing can further contribute to biological or chemical systems monitoring, new biomarkers discovery and metabolic network investigations.

Saccharomyces cerevisiae oxidative response evaluation by cyclic voltammetry and gas chromatography-mass spectrometry.

This study is focused on the evaluation of the impact of Saccharomyces cerevisiae metabolism in the profile of compounds with antioxidant capacity in a synthetic wine during fermentation. A bioanalytical pipeline, which allows for biological systems fingerprinting and sample classification by combining electrochemical features with biochemical background, is proposed. To achieve this objective, alcoholic fermentations of a minimal medium supplemented with phenolic acids were evaluated daily during 11 days, for electrochemical profile, phenolic acids, and the volatile fermentation fraction, using cyclic voltametry, high-performance liquid chromatography-diode array detection, and headspace/solid-phase microextraction/gas chromatography-mass spectrometry (target and nontarget approaches), respectively. It was found that acetic acid, 2-phenylethanol, and isoamyl acetate are compounds with a significative contribution for samples metabolic variability, and the electrochemical features demonstrated redox-potential changes throughout the alcoholic fermentations, showing at the end a similar pattern to normal wines. Moreover, S. cerevisiae had the capacity of producing chlorogenic acid in the supplemented medium fermentation from simple precursors present in the minimal medium.