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…

The role of tomato juice serum in malolactic fermentation in wine

Introduction: Malolactic fermentation (MLF) is a common process in winemaking to reduce wine acidity, maintain microbial stability and modify wine aroma. However, successful MLF is often hampered by their sluggish or stuck activity of malolactic bacteria (MLB) which may be caused by nutrient deficiency, especially when MLB are inoculated after alcoholic fermentation (Alexandre et al., 2004; Lerm et al., 2010). Identification and characterization of essential nutrients and growth factors for MLB allows for production of highly efficient nutrient supplements for MLF.

Oenological features of Sangiovese wine from vinification of whole grape berries

The present study was performed in a traditional winery located in the viticultural area of Brunello di Montalcino, Siena, Italy, in the vintage 2015. Actually, in this winery Sangiovese grape musts are fermented in large oak barrels by a single strain of Saccharomyces cerevisiae previously isolated in the same winery. Pumping over operations are carried out once or twice a day until the end of alcoholic fermentations. The aim of this work was to investigate on the oenological properties of Sangiovese wine produced with the traditional winemaking process adopted by the winery under study obtained from the fermentation of whole berries compared to that from crushed grape must. In particular, two lots of 65q of Sangiovese grapes from the same 3ha vineyard were vinified in 150hL oak barrels.

Red wine substituted esters involved in fruity aromatic expression: an enantiomeric approach to understand their sensory impact and their pathway formation

Among red wines ethyl esters, those from short hydroxylated and branched-chain aliphatic acids constitute a family with a particular behavior and sensory importance. They have been previously discussed in the literature [1] and recent studies have established that some of them were strongly involved in of red wines’ fruity aroma [2]. As some among them have an asymmetrical carbon atom, it seemed important to separate their different enantiomers to obtain an accurate assessment of their organoleptic impact. Three chiral esters have been identified, presenting alkyl and/or hydroxyle substituants: ethyl 2-hydroxy-4-methylpentanoate, ethyl 2-methylbutanoate, and ethyl 3-hydroxybutanoate.

Monitoring of Pesticide Residues from Vine to Wine

Those previous years, pesticides are often brought to the forefront by media. Questions arose about their toxicity for growers and consumers. Even if a downward trend is underway, the use of pesticides is required to ensure steady quality and quantity of harvests. A large number of active ingredients are authorized but regarding viticulture, mainly insecticides and fungicides are applied, to control pests and diseases and to increase crop yield. Some phytosanitary products, principally fungicides, applied close to the harvest date may frequently be detected in wines.

Characterizing the effects of nitrogen on grapevines with different scion/rootstock combinations: agronomic, metabolomic and transcriptomic approaches

Most vineyards are grafted and include a variety (Vitis vinifera) grafted over a wild Vitis rootstock (hybrids of V. berlandieri, riparia and rupestris). Grape berry quality at harvest depends on a subtle balance between acidity and the concentrations of sugars, polyphenols and precursors of aroma compounds. The mechanisms controlling the balance of sugars/acids/polyphenols are influenced by the abiotic environment, in particular nitrogen supply, and interact with the genotypes of both the scion variety and the rootstock. Previous work suggests that some of the effects of water stress are in fact linked to a nitrogen deficiency driven indirectly by the reduction of water absorption.