New molecular evidence of wine yeast-bacteria interaction unraveled by untargeted metabolomic profiling

The use of next-generation sequencing (NGS) has helped understand microbial genetics in oenology. Current studies mainly focus on barcoded amplicon NGS but not shotgun sequencing, which is useful for functional analyses. Since the high percentage of grapevine DNA conceals the microbial DNA in must, the majority of sequencing data is wasted in bioinformatic analyses. Here we present capture depletion of grapevine whole genome DNA.

Intra-vineyard variation grape berry ripening occurs within bunches, between bunches on the same vine and between vines. Although it is assumed that such variation also occurs at the grape berry cell wall level, no study to data has investigated in any depth. Here we have used a intra-vineyard panel design to investigate pooled bunches from six vines (per panel) in the context of a winemaking scenario. The dissected vineyard was harvested by separate panels, where each panel was then subjected to a standard winemaking procedure with or without the addition of three different enzyme preparations for maceration.

A method of suspect screening analysis to study grape metabolomics, was developed [1]. By performing ultra-high performance liquid chromatography (UHPLC) – high-resolution mass spectrometry (HRMS) analysis of the grape extract, averaging 320-450 putative grape compounds are identified which include mainly polyphenols. Identification of metabolites is performed by a new HRMS-database of putative grape and wine compounds expressly constructed (GrapeMetabolomics) which currently includes around 1,100 entries.

In the winemaking process, several compounds that remain in the grape skins and seeds after the fermentation stage are bioactive-compounds (substances with potential beneficial effects on health) that can be extracted in order to recovery valuable substances with a high commercial value for the cosmetic, food (nutraceuticals) and pharmaceutical industries. The skins contain significant amounts of bioactive substances such as tannins (16-27%) and other polyphenolic compounds (2-6.5%) in particular, catechins, anthocyanins, proanthocyanins, quercetin , ellagic acid and resveratrol.

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).