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…

An excessive leaf-fruit ratio reduces the yeast assimilable nitrogen in the must

Yeast assimilable nitrogen (YAN) in the grape must is a key variable for wine quality as a source of aroma precursors. In a situation of YAN deficiency, a foliar urea application upon the vine at veraison enhances YAN concentration and facilitates must fermentation. In 2013, Agroscope investigated the impact of leaf-fruit ratio on the nitrogen (N) assimilation and partitioning in grapevine Vitis vinifera cv. Chasselas following foliar-urea application with the aim of improving its efficiency on the YAN concentration.

The impact of different yeasts and harvest time on the wine quality of Beihong and Beimei (<I>V. vinifera x V. amurensis</I>)

Beihong and Beimei are two wine cultivars from ‘Muscat Hamberg’ (V. vinifera L.) and wild V. amurensis Rupr., which were released in China in 2008. Here,two enology practices were reported. Firstly, the impact of different yeasts including D254, GRE, K1, D21 and BDX on dry wine quality of Beihong and Beimei was investigated. For Beihong, among wines fermented by all yeasts, residual sugar content was the lowest, total anthocyanin and resveratrol contents were the highest in the wine by D254. However, the wine by D254 had lower titrable acid than those by the other yeasts except BDX.

Use of chitosan as a secondary antioxidant in juices and wines

Chitosan is a polysaccharide produced from the deacetylation of chitin extracted from crustaceous and fungi. In winemaking chitosan is mainly used in the clarification of grape juice and wine, stabilization of white wines, removal of metals and to prevent wine spoilage by undesired microorganisms. The addition of chitosan to model wine systems was able to retard browning, reduce levels of metallic ions (Fe and Cu) and to protect varietal thiols due to its antiradical activity1. The present experiment was planned in order to evaluate the use of chitosan as a secondary antioxidant at three different stages of Sauvignon blanc fermentation and winemaking. Sauvignon blanc juices from three different locations were obtained at a commercial winery in Marlborough, New Zealand. One lots of grapes was collected from a receival bin and pressed into juice with a water-bag press, and a further juice sample was collected from a commercial pressing operation. Chitosan (1 g/L, low molecular weight, 75 – 85% deacetylated) was added to the juice after pressing, after cold settling, after fermentation, or at all these stages. Controls without any chitosan additions were also prepared.

Influence of inactive dry yeast treatments during grape ripening on postharvest berry skin texture parameters and phenolic compounds extractability

Inactive dry yeast treatments in the vineyard are a tool used with the aim to improve the concentration and quality of secondary metabolites in grapes, leading to a better differentiation of the wines made from grapes differently treated. In this work, a foliar spraying treatment with yeast derivatives specifically designed to be used with the patent pending application technology of Lallemand Inc. Canada (LalVigne® Mature, Lallemand Inc., Montreal, Canada) was tested on Vitis vinifera L. cv. Barbera and Nebbiolo black winegrapes. The aim was to evaluate the effect of this treatment on the phenolic compounds accumulation, the skin physical-mechanical properties and the related phenolic extractability. Prior to analysis, the berries were sorted by flotation in order to evaluate their distribution by density class, and to determine the skin texture parameters of berries with different sugar contents, thus understanding also the ripening effect.

Defining the mechanisms and impact of winemaking treatments on tannin and polysaccharides in red wine: recent progress in creating diverse styles

Tannin and polysaccharide concentration and composition is important in defining the texture of red wines, but can vary due to factors such as cultivar, region, grape ripeness, viticultural practices and winemaking techniques. However, the concentration and composition of these macromolecules is dependent not only on grape tannin and polysaccharide concentration and composition, but also their extractability and, in the case of polysaccharides, their formation by yeast. Through studies into the influence of grape maturity, winemaking and sensory impacts of red grape polysaccharides, seed and skin tannins, recent research in our laboratory has shown that the processes involved in the extraction of these macromolecules from grapes and their retention in wine are very complex.