IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Recovery and purification of proteins from grape seed byproducts using proteomic and separative techniques

Recovery and purification of proteins from grape seed byproducts using proteomic and separative techniques

Abstract

Grape seeds account for around 5% of the weight of the whole grape berry, representing approximately 40%-50% of the solid by-products that the different wine industries generate during the winemaking process. Among all the grape seed components, proteins account for 10-13%. The use of these proteins could be of interest in technological applications for the food industry and others.
According to their solubility in different solvents, vegetable seed proteins have been classified into albumins (soluble in water), globulins (soluble in salt), prolamins (soluble in aqueous alcohol) and glutelins (soluble in acid or alkaline solution), respectively.
So far, most of the polypeptide components identified by electrophoretic analysis and mass spectroscopy in grape seed endosperm showed high homology with 11S globulin-like seed storage proteins from other plant species.
The ability of proteins to modulate food properties is highly dependent on their structural features. In this respect, there are still no studies that reveal the three-dimensional structure of these proteins in grape seed using x-ray or nuclear magnetic resonance techniques. However, there are studies using computational techniques for a 7S-type globulin from grape seed.  Therefore, the identification and subsequent elucidation of the morphology of proteins is crucial to define their potential uses and technological applications.
The aim of this work was to identify the different types of grape seed endosperm proteins from the by-product of the wine industry. For this purpose, the industrial by-product was subjected to different extractions to fractionate and purify the proteins into albumins, globulins and prolamins. In addition, quantification of the different fractions was carried out to clarify which type of protein is the majority. To carry out this work, a proteomic study based on SDS-page electrophoresis and mass spectroscopy was developed. These studies will provide new knowledge that will help to develop possible applications of seed proteins in the food industry.

DOI:

Publication date: June 23, 2022

Issue: IVAS 2022

Type: Poster

Authors

Chamizo-González Francisco1, Gordillo Arrobas Belén1, Hereida Francisco J.1, Días Ricardo2 and Freitas Víctor2

1Food Colour and Quality Laboratory, Facultad de Farmacia, Universidad de Sevilla, 41012, Sevilla, Spain
2Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade do Porto  

Contact the author

Keywords

Grape seed proteins; globulins, by-products, electrophoresis, mass spectroscopy

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

Postveraison shoot trimming in Tannat and Merlot: preliminary results on yield components, plant balance and berry composition

There is currently a trend towards the production of wines with low alcohol content. To achieve this, grapes with low sugar content must be used. There are techniques at the vineyard level that can delay ripening and avoid excessive sugar accumulation without, a priori, affecting the final polyphenol content. Postveraison shoot trimming (PVST) is experimentally evaluated for these purposes, but its impact under Uruguayan climatic conditions with high interannual variability is not known. The aim of this work is to assess the PVST in Tannat and Merlot cultivars and their impact on yield components, plant balance and berry primary composition. In this study, two commercial vineyards of 10 years old Tannat and Merlot (grafted on SO4) at Canelones Department were selected. During the 2020-201 growing season, grapevines were submitted to PVST when grapes reached 15º Brix. In a randomized block, trimmed (T) and control (C) plants were evaluated with three repetitions each cultivar. Evaluation of the evolution of primary berry composition during ripening, measurement of yield components and plant balance were performed. For both cultivars, PVST did not affect yield components. Merlot reached 5.4 kg per plant and Tannat 7.1 kg, with not statistical significance between treatments. However, statistical differences were observed in terms of plant balance. In Merlot Ravaz Index reached a difference of 5.3 (12.0 in T and 6.7 in C) meanwhile Tannat reached 3.5 of statistical difference (13.7 in T and 10.2 in C). The tendency to imbalance for the treated plants had an impact on the final grape composition. Merlot grapes showed statistical difference in final total acidity (0.3 g of difference between treatments) while treatments impact final sugar content on Tannat grapes (10.0 g of difference between treatments). Further studies are needed to assess the impact of different canopy management techniques in our conditions.

Spatial variability of temperature is linked to grape composition variability in the Saint-Emilion winegrowing area

Elevated temperature during the grape maturation period is a major threat for grape quality and thus wine quality. Therefore, characterizing the grape composition response to temperature at a larger scale would represent a crucial step towards adaptation to climate change. In response to changes in temperature, various physiological mechanisms regulate grape composition. Primary and secondary metabolisms are both involved in this response, with well-known effects, for example on anthocyanins, and lesser known effects, for example on aromas or aroma precursors. At the field scale or at the regional scale, however, numerous environmental or plant-specific factors intervene to make the effects of temperature difficult to distinguish from overall variability. In this study, it was attempted to overcome this difficulty by selecting well-characterized situations with differing temperatures.
A long-term study of air temperature variability across several Merlot vineyards in the Saint-Emilion and Pomerol wine producing area found significant temperature differences and gradients at various time scales linked to environmental factors. From this study area, a few sites were selected with similar age, soil and training system conditions, and with repeated and contrasted temperature differences during the maturation period. The average temperature difference during the maturation period was about 2°C between cooler and warmer sites, a difference similar to that expected under future climate change scenarios. In close vicinity to the temperature sensors at each site, grape berries were sampled at different times until full maturity during 2019 and 2020. Also, berries from bunches on either side of the row were analyzed separately, allowing an investigation of bunch exposure effect associated with the coupling of berry temperature and solar radiation. Four replicates of pooled berries for each time – site – bunch exposure combination were obtained and analyzed for biochemical composition. Analyses of variance of the biochemical composition data collected at different sampling times reveal significant effects associated with temperature, site, and bunch azimuth. For instance, anthocyanins in grape skins are clearly influenced by temperature and solar radiation exposure, with up to 30% reduction in warmer conditions.

Grape berry size is a key factor in determining New Zealand Pinot noir wine composition

Making high quality but affordable Pinot noir (PN) wine is challenging in most terroirs and New Zealand’s (NZ) situation is no exception. To increase the probability of making highly typical PN wines producers choose to grow grapes in cool climates on lower fertility soils while adopting labour intensive practices. Stringent yield targets and higher input costs necessarily mean that PN wine cost is high, and profitability lower, in line-priced varietal wine ranges. To understand the reasons why higher yielding vines are perceived to produce wines of lower quality we have undertaken an extensive study of PN in NZ. Since 2018, we established a network of twelve trial sites in three NZ regions to find individual vines that produced acceptable commercial yields (above 2.5kg per vine) and wines of composition comparable to “Icon” labels. Approximately 20% of 660 grape lots (N = 135) were selected from within a narrow juice Total Soluble Solids (TSS) range and made into single vine wines under controlled conditions. Principal Component Analysis of the vine, berry, juice and wine parameters from three vintages found grape berry mass to be most effective clustering variable. As berry mass category decreased there was a systematic increase in the probability of higher berry red colour and total phenolics with a parallel increase in wine phenolics, changed aroma fraction and decreased juice amino acids. The influence of berry size on wine composition would appear stronger than the individual effects of vintage, region, vineyard or vine yield. Our observations support the hypothesis that it is possible to produce PN wines that fall within an “Icon” benchmark composition range at yields above 2.5kg per vine provided that the Leaf Area:Fruit Weight ratio is above 12cm2 per g, mean berry mass is below 1.2g and juice TSS is above 22°Brix.

Assessing the relationship between cordon strangulation, dieback, and fungal trunk disease symptom expression

Grapevine trunk diseases including Eutypa dieback are a major factor in the decline of vineyards and may lead to loss of productivity, reduced income, and premature reworking or replanting. Several studies have yielded results indicating that vines may be more likely to express symptoms of vascular disease if their health is already compromised by stress. In Australia and many other wine-growing regions it is a common practice for canes to be wrapped tightly around the cordon wire during the establishment of permanent cordon arms. It is likely that this practice may have a negative effect on health and longevity, as older cordons that have been trained in this manner often display signs of decay and dieback, with the wire often visibly embedded within the wood of the cordon. It is possible that adopting a training method which avoids constriction of the vasculature of the cordon may help to limit the onset of vascular disease symptom expression. A survey was conducted during the spring of two consecutive growing seasons on vineyards in South Australia displaying symptoms of Eutypa lata infection when symptomless shoots were 50–100 cm long. Vines were assessed as follows: (i) the proportion of cordon exhibiting dieback was rated using a 0–100% scale; (ii) the proportion of canopy exhibiting foliar symptoms of Eutypa dieback was rated using a 0–100% scale; (iii) the severity of strangulation was rated using a 0–4 point scale. Images were also taken of each vine for the purpose of measuring plant area index (PAI) using the VitiCanopy App. The goal of the survey was to determine if and to what extent any correlation exists between severity of strangulation and cordon dieback, in addition to Eutypa dieback foliar symptom expression.

Influence of a spontaneous cover crop on the vineyard and soil erosion under Mediterranean climate

Sixty five % of the agricultural area of the Basque Country located in the DO Ca Rioja corresponds to vineyards. More than 40% of it has an average slope greater than 10%, which makes it sensitive to erosive processes. Furthermore, it is foreseeable that extreme weather events (storms, hail, extreme heat and cold, etc.) will be favored due to climate change. Cover cropping can mitigate this risk, and therefore the objective of this work is to evaluate the impact that a vegetable cover has on the agronomic behavior of the vineyard, the quality of the grape and soil erosion. For this, a trial has been carried out with a Graciano variety vineyard with a slope between 10% -20% during the years 2020 and 2021. Conventional tillage management in the area has been compared (4-6 passes per year of tillage machinery) versus spontaneous vegetation cover management in the vineyard. This implies not tilling and allowing the grass of the land to colonize the range between the lines of vines, controlling their height through 1-3 mowing passes per year, always trying to affect the surface of the land as little as possible. The vegetative growth, yield and quality of the grape and wine was measured. Furthermore, erosion has been measured using Gerlasch boxes. The yield was lower in the second year of the trial in the cover crop treatment, but erosion was significantly reduced.