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…

Simultaneous monitoring of dissolved CO2 and collar from Rosé sparkling wine glasses: the impact of yeast macromolecules

Champagne or sparkling wines elaborated through the same traditional method, which consists in two major yeast-fermented steps, typically hold about 10 to 12 g/L of dissolved CO2 after the second fermentation in a closed bottle. Hundreds of molecules and macromolecules originating from grape and yeast cohabit with dissolved CO2; they are essential compounds contributing to many organoleptic characteristics (effervescence, foam, aroma, taste, colour…). Indeed, the second alcoholic fermentation and the maturation on lees (which may last from 12 months up to several years) both induce various quantitative and qualitative changes in the wine through the action of yeast, as listed hereafter: development of aromas during aging on lees, release of nitrogen compounds during autolysis and release of macromolecules (polysaccharides, lipids, nucleic acids) in wine.

Extraction of pathogenesis-related proteins and phenolics in Sauvignon Blanc as affected by different

The composition of wine is largely determined by the composition of pre-fermentation juice, which is influenced by extraction of grape components. Different grape harvesting and processing conditions could affect the extraction of grape components into juice. Among these grape components, pathogenesis-related (PR) proteins are of great concern for white wine maker as they are the main cause of haze formation in finished white wine. If not removed before bottling, these PR proteins may progress into haze through the formation of complex with phenolics under certain conditions. Thaumatin-like proteins (TLPs) and chitinases are the main constituents of PR proteins found in protein haze.

Characterization of various groups of pyranoanthocyanins in Merlot red wine

In red wines, anthocyanins evolve during the wine-making process and ageing. They react with other compounds (such as vinylphenols, acetaldehyde, pyruvic acid…) to form a stable family of compounds called pyranoanthocyanins. Furthermore, the oxidation process can modify the anthocyanic profile of a red wine. It is also interesting to evaluate the occurrence of the different subclasses of pyranoanthocyanins and to characterize their chemical properties. The first objective of this study is to evaluate the occurrence of the different groups of pyranoanthocyanins in an oxidised Merlot wine by a centrifugal partition chromatography strategy. The second goal is to evaluate their relative impact in red wines from Bordeaux region by measuring their concentrations.

Grape byproducts as source of resveratrol oligomers for the development of antifungal extracts

Grape canes are a non-recycled byproduct of wine industry (1-5 tons per hectare per year) containing valuable phytochemicals of medicine and agronomical interest. Resveratrol and wine polyphenols are known to exert a plethora of health-promoting effects including antioxidant capacity, cardioprotection, anticancer activity, anti-inflammatory effects, and estrogenic/antiestrogenic properties (Guerrero et al. 2009). Additionally, resveratrol is a major phytoalexin produced by plants in response to various stresses and promotes disease resistance (Chang et al. 2011). Our project aims to develop polyphenol-rich grape cane extracts to fight phytopathogenic or clinically relevant fungi. We initiate the project with the development of analytical methods to analyze resveratrol mono- and oligomers (dimers, trimers and tetramers) from grape canes and we evaluate their potential activity against clinically relevant opportunistic fungal pathogens (Houillé et al. 2014).

A preliminary study of clonal selection in cv. Viura in relation to varietal aroma profile

Viura is a synonym for Macabeo and currently it is the most widely planted white grape variety in D.O.Ca. Rioja, with 3,569 ha, representing 84% of the white grape cultivated area. It is a generous-yielding grape, presenting low values of titratable acidity and with large and compact clusters which makes it susceptible to Botrytis cinerea. Thus, this variety not always satisfies the wine grower’s prospects. Nowadays, the available plant material is scarce, moreover, it was selected on the basis of other quality criteria, not currently requested.