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…

Modulating role of SO2 in white wine protein haze formation

Despite the extensive research performed during the last decades, the multifactorial mechanism responsible for the white wine protein haze formation is not fully characterized. Herein, a new model is proposed, which is based on the experimental identification of sulfur dioxide as a major modulating factor inducing wine protein haze upon heating. As opposed to other reducing agents, such as 2-mercaptoethanol, dithiothreitol and tris(2-carboxyethyl)phosphine hydrochloride (TCEP), the addition of SO2 to must/wine upon heating cleaves intraprotein disulfide bonds, hinders thiol-disulfide exchange during protein interactions and can lead to the formation of novel inter/intraprotein disulfide bonds. Those are eventually responsible for wine protein aggregation which follows a nucleation-growth kinetic model as shown by dynamic light scattering [1].

Interest and impact of PVP/PVI (Polyvinylpyrrolidone/ Polyvinylimidazole) on winemaking and final quality of wines

Céline Sparrow a, Christophe Morge a, a SOFRALAB SAS, 79, av. A.A. Thévenet – CS 11031 – 51530 Magenta, France Consumers’ health and security force authorities to limit, in wine as in others food industry products, the concentration in « dangerous » molecules. Therefore the legal limit in heavy metals keeps on decreasing. As per proof EU regulation just decrease the stain concentration in wine from 0,2 to 0,15 mg/l. Certain changes , such as sodium arsenite treatment in vines, disappearance of brass in wineries to the benefit of stainless steel, limit even more the concentration of heavy metals in wines. But the use of copper derivates in vines treatments is difficult to replace. In the case of wine and its elaboration, the problem is even more complex. Indeed, regulation forces the wine producers to control the concentration of certain heavy metals in final wines.

Ripening of cv. Cabernet Sauvignon grapes: polysaccharides fractions evolution and phenolic extractability

Polysaccharides and more specifically pectins, make up a significant portion of the cell wall material of the plant cells including the grapes. During the fruit ripening the associated softening is related to the breakdown of the cell wall polysaccharides. During this process, it is expected that polysaccharides that are soluble in red wine will be formed influencing its texture. Anthocyanins are responsible for the wine color and tannins for the astringency, body and bitterness of the wine. In the skins, these compounds are located in the cell vacuoles and the barrier that conditions their extractability is the skin cell wall that may determine the mechanical resistance, the texture and the ease of processing berries. The aim of this work was study the evolution of the polysaccharides and the anthocyanin and tannin extractability during the ripening period in Cabernet Sauvignon grapes, trying to correlate these variables.

New biological tools to control and secure malolactic fermentation in high pH wines

Originally, the role of the malolactic fermentation (MLF) was simply to improve the microbial stability of wine via biological deacidification. However, there is an accumulation of evidence to support the fact that lactic acid bacteria (LAB) also contribute positively to the taste and aroma of wine. Many different LAB enter into grape juice and wine from the surface of grape berries, cluster stems, vine leaves, soil and winery equipment. Due to the highly selective environment of juices and wine, only a few types of LAB are able to grow.

Contribution of Piperitone to the mint nuances perceived in the aging bouquet of red Bordeaux wines

During the tasting of a fine, old wine, the aromas generated in the glass are intertwined in an intimate, complex manner, expressing the fragrance of the aging bouquet. This aging bouquet, which develops during bottle storage through a complex transformation process, may result in a broad palette of nuances. Among these, undergrowth, truffle, toasted, spicy, licorice, fresh red- and black-berry fruit and mint descriptors were recently identified as features of its olfactory representation for red Bordeaux wines. Although a targeted chemical approach focusing on volatile sulfur compounds revealed the role played by dimethyl sulfide, 2-furanmethanethiol, and 3-sulfanylhexanol as molecular markers of the typicality of the wine aging bouquet of red Bordeaux wines, its chemical transcription has only partially been elucidated.