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…

New acylated flavonols identified in the grape skin of Vitis vinifera cv. Tannat and their wines

Flavonols are a class of flavonoid compounds derived from plant secondary metabolism. There they play different roles like antioxidants, internal regulators and UV screenings. In red wines, flavonols have increasingly received consideration by part of scientific and winemakers according their properties began to arise known. Among these stand out wine colour stabilization and their value as bioactive compounds. In this work the complete series of the acetylated and p-coumaroylated derivatives of the 3-O-glycosides of methoxylated flavonols, namely isorhamnetin, laricitrin and syringetin, have been identified in grapes and their respective wines from Vitis vinifera cv. Tannat.

Innovations in the use of bentonite in enology: interactions with grape and wine proteins, colloids, polyphenols and aroma compounds.

The use of bentonite in oenology rounds around the limpidity and the stability that determine consumer acceptability. As a matter of fact, the haze formation in wine reduces its commercial value and makes it unacceptable for sale. Stabilization treatments are, therefore, essential to ensure a long-time limpidity and to forecast the formation of deposits in the bottle. Bentonite that is normally used in oenology for clarifying-fining purpose, shows a natural clay-based mineral structure allowing it to swell and to jelly in water and hence in must and wine.

Influence of preflowering basal leaf removal on aromatic composition of cv. Tempranillo wine from semiarid climate (Extremadura Western Spain)

Abstract In this work the effects of early leaf removal performed manually at preflowering phenological stage, on the volatile composition of Tempranillo (Vitis vinifera L.) wines were studied. From 2009-2011 vintages 34 wine volatile compounds were identified and quantified by gas chromatography-mass spectrometry (GC-MS) where early leaf removal only modified 25 of them. The total C6 compounds, acetates and volatiles acids (with exception of isobutyric acid) were affected by defoliation, whereas alcohols and esters showed a minor effect. Furthermore the vintage effect also was shown.

Impact of glutathione and elemental sulphur juice addition on the volatile thiol production in South African Sauvignon blanc wine

Three compounds, 3-mercaptohexanol (3MH), 3-mercaptohexyl-acetate (3MHA) and 4-mercapto-4-methylpentan-2-one (4MMP), also known as varietal thiols, have been identified to contribute positively to wine aroma and are responsible for the distinct gooseberry, grapefruit, guava and box tree character found in Sauvignon blanc wines. Certain volatile thiol compounds though, can cause off-aromas of onion, garlic, rubber and rotten egg, this group of molecules is known as reductive sulphur compounds (RSC). This study looks into how the addition of sulphur-compounds to Sauvignon blanc juice contributes to the varietal thiol (3MH and 3MHA) concentration and reductive sulphur compound concentration in South African Sauvignon blanc wine.

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.