Macrowine 2021
IVES 9 IVES Conference Series 9 Some applications come from a method to concentrate proteins

Some applications come from a method to concentrate proteins

Abstract

All techniques usually used to assay proteins was not reliable in vegetable extract due to interferences with the components included in extracts like polyphenols, tanins, pectines, aromatics compounds. Absorbance at 280nm, Kjeldhal assay, Biuret and Lowry methods, Acid Bicinchonique technique and Bradford assay give the results depending on the composition of extract, on the presence or not of detergent and on the raw material (Marchal, 1995). Another difficulty in these extracts for the quantification of proteins comes from the large amount of water included in vegetable and the low concentration of proteins. Thus in red wines, proteins are usually not taken into account due to their low concentration (typically below 10 mgL-1) and to the presence of anthocyanis and polyphenols. Due to all these defects, alternative test should be developed. The perfect protein assay would exhibit the following characteristics: fast, easy to use, sensitive, accurate, precice and free from interferences. Futhermore this assay should be compatible with all substances commonly found in protein samples and at low concentration. Our purpose in this work is to combine the concentration of proteins by bentonite with separation electrophoretic 1D SDS Page and to examine some applications. First, wines were fined with 100g/hl of bentonite is largely sufficient to adsorbe all proteins (Paetzold and al.,1990). In these conditions, we observed at low concentrations of bentonite (under 20g/hl), the bentonite Electra® adsorbed only β glucanases and chitinases. Second after desorption by Laemmli buffer, proteins were separated by SDS-PAGE and quantified after coloration with Coomassie Blue R-250 by scanning coupled to the image analysis TotalLab software (Sauvage and al., 2010). The gels after destaining were scanned with a transmission scanner at 300 dpi to obtain a digitised image. The software compared the volume (area x intensity of each pixel) of each band to the volume of BSA band (included in standard file). Each band was characterized by the molecular weight and the quantity of proteins expressed in µg equivalent BSA. The sum of each band gave the total pool of proteins included in each sample. The standard deviation measured on 6 same sample on Chardonnay wine was 11%. The response was linear for each band up to 1µg/band. By this method we also got the relative composition of the majority of proteins. Last, but not least, proteins were desorbed from bentonite with buffer to denature proteins or only with a buffer to conserve the native form of proteins (like Tris buffer or NaCl solution). After this experimentation we checked if [1] Marchal R. Ph. Thesis, university of Reims, 1995.

[1] Paetzold M., Dulau L., Dubourdieu D. J.Inter.Sci.Vigne Vin, 1990, 24, 13-28. [2] Pocock K.F., Waters E.J. Aust.J.Grape Wine Res., 4, 136-139. [4] Sauvage F.X., Bach B., Moutounet M., Vernhet A. 2009, 118, 26-34.

Publication date: May 17, 2024

Issue: Macrowine 2016

Type: Poster

Authors

Francois-Xavier Sauvage*, Patrick Chemardin

*INRA

Contact the author

Tags

IVES Conference Series | Macrowine | Macrowine 2016

Citation

Related articles…

Chemical markers in wine related to low levels of yeast available nitrogen in the grape

Nitrogen is an important nutrient of yeast and its low content in grape must is a major cause for sluggish fermentations. To prevent problems during fermentation, a supplementation of the must with ammonium salts or more complex nitrogen mixtures is practiced in the cellar. However this correction seems to improve only partially the quality of wine [1]. In fact, yeast is using nitrogen in many of its metabolic pathways and depending of the sort of the nitrogen source (ammonium or amino acids) it produces different flavor active compounds. A limitation in amino acids can lead to a change in the metabolic pathways of yeast and consequently alter wine quality.

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.

Effect of nanofiltration on the chemical composition and wine quality

In Enology the conventional processes of filtration for clarification and stabilization are giving place to alternative membrane processes, including nanofiltration (NF). Furthermore, the increased alcohol content in wines recorded in recent years became an important issue for all the main wine producing countries. Among techniques available to the wine industry to reduce the ethanol content, NF is certainly one of the newest. This study is focused on the evaluation of NF influence on wine physical-chemical composition, including mineral content, which in accordance to our best knowledge is a novelty.

Directed Evolution of Oenococcus oeni: optimising yeast-bacteria interactions for improved malolactic fermentation

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.

Effect of supplementation with inactive yeast during alcoholic fermentation in base wine for sparkling

INTRODUCTION: Foam stability of sparkling wines is significantly favored by the presence of surface active agents such as proteins and polysaccharides [1]. For that reason, the renowned sparkling wines are aged after the second fermentation in contact with the lees for several months (even years). Thereby wines are enriched in these macromolecules due to yeast autolysis. Since this practice is slow and costly, winemakers are seeking for alternative procedures to increase their concentration in base wines. In that sense, the supplementation with inactive yeast during alcoholic fermentation has been proposed [2]. The aim of this study was to determine whether this new strategy is really useful for enriching base wines in macromolecules and for improving foam properties of the base wines.