Proteomic and activity characterization of exocellular laccases from three Botrytis cinerea strains

Consumers predominantly use visual, aromatic and texture cues as quality/preference indicators to describe olfactory sensations. In this study, the effect of micro-organism in wine production was investigated using analytical and sensory techniques to achieve relevant analytical characterisation. Selected anthocyanins, flavan-3-ols, flavonols and phenolic acids were quantified in Syrah wines using RP-HPLC-DAD. Standard oenological parameters were also measured. Syrah grape must was fermented with various combinations of Saccharomyces cerevisiae (S. cerevisiae) and non-Saccharomyces (Metschnikowia pulcherrima or Hanseniaspora uvarum) yeasts, which was followed by sequential inoculation of lactic acid bacteria (LAB) (Oenococcus oeni or Lactobacillus plantarum).

Introduction: Malolactic fermentation (MLF) is a common process in winemaking to reduce wine acidity, maintain microbial stability and modify wine aroma. However, successful MLF is often hampered by their sluggish or stuck activity of malolactic bacteria (MLB) which may be caused by nutrient deficiency, especially when MLB are inoculated after alcoholic fermentation (Alexandre et al., 2004; Lerm et al., 2010). Identification and characterization of essential nutrients and growth factors for MLB allows for production of highly efficient nutrient supplements for MLF.

The occurrence of faults and taints in wine, such as those caused by microbial spoilage or various taints, have resulted in significant financial losses to wine producers. The wine industry commits significant financial resources towards fining and taint removal processes each year. Fining involves the addition of one or more adsorptive substrates to juice or wine to bind certain components, thus reducing their concentration [1]. However, these processes are often not selective and can also remove desirable flavour and aroma compounds.

Despite the extensive research performed during the last decades, the multifactorial mechanism responsible for the white wine protein haze formation is not fully characterized. Herein, a new model is proposed, which is based on the experimental identification of sulfur dioxide as a major modulating factor inducing wine protein haze upon heating. As opposed to other reducing agents, such as 2-mercaptoethanol, dithiothreitol and tris(2-carboxyethyl)phosphine hydrochloride (TCEP), the addition of SO2 to must/wine upon heating cleaves intraprotein disulfide bonds, hinders thiol-disulfide exchange during protein interactions and can lead to the formation of novel inter/intraprotein disulfide bonds. Those are eventually responsible for wine protein aggregation which follows a nucleation-growth kinetic model as shown by dynamic light scattering [1].

For characterization, sensory designing and authentication rapid analytical technologies have become available. Some, like Proton Transfer Reaction Mass Spectrometry allow a rapid spectrum of the volatile compounds of wines. Combined with chemometrics wines can be characterized. The same approach can be used to calculate the results of virtual mixtures and allow formulation of constant quality blends. Other new techniques and portable devices based on spectroscopy allow measurements on production sites and in grocery stores, even for the smart consumer. We will present some examples of the application of these techniques for authentication of wines, both in the laboratory and on site.