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…

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 malolactic fermentation in barrels or stainless steel tanks on wine composition. Influence of the barrel toasting

Ellagitannin, anthocyanin and woody volatile composition of Cabernet Sauvignon wines aged in oak barrels for 12 months was evaluated. Depending on the container where malolactic fermentation (MLF) was carried out, two wine modalities were investigated: wines with MLF carried out in stainless steel tanks and barrel-fermented wines. Three toasting methods (medium toast, MT; medium toast with watering, MTAA; noisette) were considered for ageing of each wine modality. Sensory analyses (triangle and rating tests) were also performed. Two-way ANOVA of the raw experimental data revealed that the toasting method and the container where MLF took place, as well as the interaction between both factors, have a significant influence (p < 0.05) on ellagitannin, anthocyanin and woody volatile profiles of Cabernet Sauvignon wines.

Impact of some agronomic practices on grape skins anthocyanin content

Wine colour is the first quality characteristic to be assessed, especially regarding red wines. Anthocyanins are very well known to be the main responsible compounds for red wine colour. Red cultivars can synthesize and accumulate anthocyanins in berry skin to express their colour. However, anthocyanin accumulation is often influenced by a series of factors, such as genetic regulation, phytohormones, environmental conditions and viticultural management.

Determination of metallic elements in Chilean wines by atomic absorption spectroscopy and inductively coupled plasma–mass spectrometry

The chemical composition of wines depends on series of variables such as the type of grape, edaphoclimatic conditions, and viticulture and winemaking practices employed during production. Metallic elements play a significant role during winemaking (e.g. as catalysts of oxidation reactions) and have been previously employed for the classification of wines according to provenance. In this work, we focused on the analysis of metallic elements (K, Na, Ca, Zn, Cu, Fe, Mg, Mn, Ni, Cr, Al, Pb, Cd, Hg, Se, Co, Sn and As) in 145 Chilean wine samples (102 reds and 43 white wines), of seven grape varieties, and five of the major wine producing regions in Chile.

Impact of smoke exposure on the chemical composition of grapes

Vineyard exposure to smoke can lead to grapes and wine which exhibit objectionable smoky and ashy aromas and flavours, more commonly known as ‘smoke taint’ [1, 2]. In the last decade, significant bushfires have occurred around the world, including near wine regions in Australia, Canada, South Africa and the USA, as a consequence of the warmer, drier conditions associated with climate change. Considerable research has subsequently been undertaken to determine the chemical, sensory and physiological consequences of grapevine exposure to smoke. The sensory attributes associated with smoke-tainted wine have been linked to the presence of several smoke-derived volatile phenols, such as guaiacols, syringols and cresols [2].