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…

Interactions of wine polyphenols with dead or living Saccharomyces cerevisiae Yeast Cells and Cell Walls: polyphenol location by microscopy

Tannin, anthocyanins and their reaction products play a major role in the quality of red wines. They contribute to their sensory characteristics, particularly colour and astringency. Grape tannins and anthocyanins are extracted during red wine fermentation. However, their concentration and composition change over time, due to their strong chemical reactivity1. It is also well known that yeasts influence the wine phenolic content, either through the release of metabolites involved in the formation of derived pigments1, or through polyphenol adsorption2,3.

Effect of the winemaking technology on the phenolic compounds, foam parameters in sparklig wines

Contribution Sparkling wines elaborated following the traditional method undergo a second fermentation in closed bottles of base wines, followed by aging of wines with lees for at least 9 months. Most of the sparkling wines elaborated are white and rosé ones, although the production of red ones is highly increasing. One of the initial problems in red sparkling wine processing is to obtain suitable base wines that should have moderate alcohol content and astringency and adequate color intensity; which is difficult to obtain when grapes must be harvested at low phenolic and industrial maturity stage. The low phenolic maturity degree in the red grapes makes essential to choose an adequate winemaking methodology to obtain the base wines because the extracted polyphenols will vary according the winemaking technique: carbonic maceration or destemmed-crushed grapes.

Mean polymerization degree of proanthocyanidins of grape seeds, skins and wines from Agiorgitiko (cv. Vitis vinifera): Differences among vintages

Grape phenolic compounds are very important constituents of red wine because, in addition to their antioxidant properties, they contribute to color, astringency and bitterness, oxidation reactions, interactions with proteins and ageing behavior of wines. The aim of our study was to assess the structural characteristics of grape and wine proanthocyanidins of Agiorgitiko variety and to evaluate the influence of the vintage year. Twelve vineyard locations were designated in the Nemea wine region. For three consecutive years (2012-2014), the grapes were harvested at technological maturity and the method of phloroglucinolysis was employed to determine the mean degree of polymerization (mDP) and subunit composition of the samples.

Fingerprinting the origin of rosé wines with a new high throughput polyphenomics method

Wine is a widely consumed alcoholic beverage with a high commercial value. More specifically, the worldwide consumption of rosé wine has increased by 20% since 2002[1]. But because of its high commercial value, it can become a subject of fraud, and authenticity control is necessarily required. More than one hundred polyphenols have been recently quantified in various rosé wines [2]. They are key components defining color, taste and quality of wines. Their amount and composition depend on many different factors such as grape variety, winemaking and age of the wine. In this study, the influence of geographic origin of some rosé French wines was investigated. An original and very fast UPLC-QTOF-MS method was developed and used to predict the geographic origin authenticity of rosé wines.

To a better understanding of the impact of vine nitrogen status on volatile thiols from plot to transcriptome level

Volatile thiols contribute largely to the organoleptic characteristics and typicity of Sauvignon blanc wines. Among this family of odorous compounds, 3-sulfanylhexan-1-ol (3SH) and 4-methyl-4-sulfanylpentan-2-one (4MSP) have a major impact on wine flavor. These thiols are formed during alcoholic fermentation by the yeast from odorless and non-volatile precursors found in the berry and the must. The effect of vine nitrogen status on 3SH and 4MSP in Sauvignon blanc wine and on the glutathionylated and cysteinylated precursors of 3SH (Glut-3SH and Cys-3SH) was investigated in this study.