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…

On the losses of dissolved CO2 during champagne aging

A misconception lingers in the minds of some wine consumers that Champagne wines don’t age. It’s largely a myth, certainly as far as the best cuvees are concerned. Actually, during the so-called autolysis period of time (in the closed bottle, after the “prise de mousse”), complex chemical reactions take place when the wine remains in contact with the dead yeast cells, which progressively bring complex and very much sought-after aromas to champagne. Nevertheless, despite their remarkable impermeability to liquid and air, caps or natural cork stoppers used to cork the bottles are not 100% hermetic with regard to gas transfers. Gas species therefore very slowly diffuse through the cap or cork stopper, along their respective inverse partial pressure. After the “prise de mousse”, because the partial pressure of CO2 in the bottleneck reaches up to 6 bars (at 12 °C), gaseous CO2 progressively diffuse from the bottle to the ambient air
(where the partial pressure of gaseous CO2 is only of order of 0,0004 bar).

The impact of branched chain and aromatic amino acids on fermentation kinetics and aroma biosynthesis by wine yeast Saccharomyces cerevisiae

One of the major determinants of wine quality is the aroma. Wine aroma is the human perception of the matrix of grape and yeast derived volatiles and their interaction that contribute to flavour wine. Most common are higher alcohols, ester and aldehydes. In previous studies the formation of characteristic volatile compounds have been linked to the metabolism of branched-chain and aromatic amino acids
(BCAAs) in synthetic grape must. Here we report on an investigation to assess the impact of the initial amino acid concentration on the production of aroma compounds by the industrial yeast VIN13 grown in both synthetic and real grape musts.

Impact of varying ethanol and carbonation levels on the odor threshold of 1,1,6-trimethyl-1,2-dihydronaphtalene (petrol off-flavor) and role of berry size and Riesling clones

1,1,6-trimethyl-1,2-dihydronaphtelene (TDN) evokes the odor of “petrol” in wine, especially in the variety Riesling. Increasing UV-radiation due to climate change intensifies formation of carotenoids in the berry skins and an increase of TDN-precursors1. Exploring new viticultural and oenological strategies to limit TDN formation in the future requires precise knowledge of TDN thresholds in different matrices. Thresholds reported in the literature vary substantially between 2 µg/L up to 20 µg/L2,3,4 due to the use of different methods. As Riesling grapes are used for very different wine styles such as dry, sweet or sparkling wines, it is essential to study the impact of varying ethanol and carbonation levels.

Impact of drought stress on concentration and composition of wine proteins in Riesling

Protein haze in white wines is a major technological and economic problem of the wine industry. Field tests were carried out in steep slope vineyards planted with Riesling grapes over 3 dry growing seasons to study the effect of drought stress on the concentration of proteins in the resulting wines. Plots suffering from drought stress were compared with surrounding drip irrigated plots. Riesling grapes were processed into wines by conventional procedures. Protein amounts of the isolated wine colloids of the stressed samples were always higher than those of the watered samples(mean watered 13.8 ± 0.44, mean stressed 17.4 ± 0.40 g 100 g-1). As a consequence, higher bentonite doses were needed to achieve protein haze stability of the drought stressed treatments.

Assessing the effect of oak derived aromas on mouthfeel perception in Chardonnay wine

Mouthfeel is an important quality parameter for Chardonnay wines, particularly those aged in oak. While research on mouthfeel has traditionally focused on the impact of non-aromatic compounds, the role of aroma compounds has largely been over looked. However, in wine as well as other food interactions between retronasal aroma and mouthfeel have been noted. The goal of this research was to investigate the impact of wine aroma on the perception of mouthfeel. Because of the importance of oak aging in the development of Chardonnay mouthfeel, the impact of oak aromas on perceived mouthfeel was explored. Aroma compounds associated with oak (ethyl palmitate, eugenol, furfural, isoeugenol, syringaldehyde, vanillin and whiskey lactone) were added to two different Chardonnay wines; one with no oak influence and one fermented in neutral oak. Low and high concentrations of the compounds were added based on concentrations typically found in barrel aged Chardonnay wine.