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…

Phenolic composition of Tempranillo Blanco grapes changes after foliar application of urea

Our research aimed to determine the effect and efficiency of foliar application of urea on the phenolic composition of Tempranillo Blanco grapes. The field experiment was carried out in 2019 and 2020 seasons and the plot was located in D.O.Ca Rioja (North of Spain). The vineyard was Vitis vinifera L. Tempranillo Blanco and grafted on Richter-110 rootstock. The treatments were control (C), whose plants were sprayed with water and three doses of urea: plants were sprayed with urea 3 kg N/ha (U3), 6 kg N/ha (U6) and 9 kg N/ha (U9). The applications were performed in two phenological stages, pre-veraison (Pre) and veraison (Ver). Also, each of the treatments was repeated one week later. Control and treatments were performed in triplicate and arranged in a randomised block design. Grapes were harvested at optimum ripening stage. High-performance liquid chromatography was used to analyse the phenolic composition of the grapes. Finally, the results obtained from the analytical determinations – flavonols, flavanols and non-flavonoid (hydroxybenzoic acids, hydroxycinnamic acids and stilbenes) – were studied statistically by analysis of variance. The results showed that, in 2019, U6-Pre and U9-Pre treatments increased the hydroxybenzoic acid content in grapes, and also all foliar treatments applied at Pre enhanced the stilbene concentration. Moreover, U3-Ver was the only treatment that rose flavonol and stilbene contents in the Tempranillo Blanco grapes. In 2020, all treatments applied at Pre enhanced the flavonol concentration in grapes. Furthermore, U3-Pre and U9-Pre treatments increased stilbene content in grapes. Nevertheless, the hydroxybenzoic acid content was improved by U6-Ver and U9-Ver and besides, hydroxycinnamic acid concentration in grapes was increased by all treatments applied at Ver. In conclusion, the lower and highest dose of urea (U3 and U9), applied at pre-veraison, were the best treatments to improve the Tempranillo Blanco grape phenolic composition.

A multidisciplinary approach to evaluate the effects of the training system on the performance of “Aglianico del Vulture” vineyards

Vineyards are complex agro-ecosystems with high spatial and temporal variability. An efficient training system may counteract the adverse effects of this variability. Moreover, considering the climate change issues, choosing an efficient training system that enhances water use and protects the vines from radiative thermal stress has become a priority for the farmers. A multidisciplinary approach that assesses the soil-crop-yield-wine relationships of vineyards in a distributed and holistic way could bring added knowledge on the behavior of the different training systems. This ongoing research aimed to implement a multidisciplinary approach to study the behavior of “Aglianico del Vulture” grapevines trained with two different systems: a spurred cordon (SC) and an “Alberello in parete” (AL), grown in a high-quality wine production area of Basilicata region (Italy). The approach merged several methods and scales of soil, ecophysiology, must/wine quality, and spectral data collection to assess the influence of the training system. Homogeneous zones (HZs) in both training systems were defined through a procedure based on geomorphological classification, unmanned aerial vehicles (UAV) images analysis, and a traditional soil survey supported by geophysical scanning. During the 2021 season, TDR probes monitored soil water content, while grapevine health status was assessed using eco-physiological measurements (LWP, chlorophyll content, PSII photosynthetic efficiency, LAI, and point-based field spectroscopy). These grapevine in-vivo measurements validated the spectral vegetation indexes (NDVI, RENDVI, CVI, and TVI) derived from the UAV multispectral imagery, which monitored the grapevine status in a distributed and non-invasive way. Grape yield, quality of berries, must and wine were measured to assess the effects of the training systems. The first experimental year results showed the variability of the vineyards and revealed relationships among soil parameters, crop characteristics, and vegetation indices of the SC and AL training systems. This multidisciplinary study could bring new insights into the vineyard training system’s effects on grape yield and wine quality.

Elevational range shifts of mountain vineyards: Recent dynamics in response to a warming climate

Increasing temperatures worldwide are expected to cause a change in spatial distribution of plant species along elevational gradients and there are already observable shifts to higher elevations as a consequence of climate change for many species. Not only naturally growing plants, but also agricultural cultivations are subject to the effects of climate change, as the type of cultivation and the economic viability depends largely on the prevailing climatic conditions. A shift to higher elevations therefore represents a viable adaptation strategy to climate change, as higher elevations are characterized by lower temperatures. This is especially important in the case of viticulture because a certain wine-style can only be achieved under very specific climatic conditions. Although there are several studies investigating climatic suitability within winegrowing regions or longitudinal shifts of winegrowing areas, little is known about how fast vineyards move to higher elevations, which may represent a viable strategy for winegrowers to maintain growing conditions and thus wine-style, despite the effects of climate change. We therefore investigated the change in the spatial distribution of vineyards along an elevational gradient over the past 20 years in the mountainous wine-growing region of Alto Adige (Italy). A dataset containing information about location and planting year of more than 26000 vineyard parcels and 30 varieties was used to perform this analysis. Preliminary results suggest that there has been a shift to higher elevations for vineyards in general (from formerly 700m to currently 850 m a.s.l., with extreme sites reaching 1200 m a.s.l.), but also that this development has not been uniform across different varieties and products (i.e. vitis vinifera vs hybrid varieties and still vssparkling wines). This is important for climate change adaptation as well as for rural development. Mountain areas, especially at mid to high elevations, are often characterized by severe land abandonment which can be avoided to some degree if economically viable and sustainable land management strategies are available.

Influence of grapevine rootstock/scion combination on rhizosphere and root endophytic microbiomes

Soil is a reservoir of microorganisms playing important roles in biogeochemical cycles and interacting with plants whether in the rhizosphere or in the root endosphere. The composition of the microbial communities thus impacts the plant health. Rhizodeposits (such as sugar, organic and amino acids, secondary metabolites, dead root cells …) are released by the roots and influence the communities of rhizospheric microorganisms, acting as signaling compounds or carbon sources for microbes. The composition of root exudates varies depending on several factors including genotypes. As most of the cultivated grapevines worldwide are grafted plants, the aim of this study was to explore the influence of rootstock and scion genotypes on the microbial communities of the rhizosphere and the root endosphere. The work was conducted in the GreffAdapt plot (55 rootstocks x 5 scions), in which the 275 combinations have been planted into 3 blocks designed according to the soil resistivity. Samples of roots and rhizosphere of 10 scion x rootstock combinations were first collected in May among the blocks 2 and 3. The quantities of bacteria, fungi and archaea have been assessed in the rhizosphere by quantitative PCR, and by cultivable methods for bacteria and fungi. The communities of bacteria, fungi and arbuscular mycorrhizal fungi (AMF) was analyzed by Illumina sequencing of 16S rRNA gene, ITS and 28S rRNA gene, respectively. The level of mycorrhization was also evaluated using black ink coloration of newly formed roots harvested in October. The level of bacteria, fungi and archaea was dependent on rootstock and scion genotypes. A block effect was observed, suggesting that the soil characteristics strongly influenced the microorganisms from the rhizosphere and root endosphere. High-throughput sequencing of the different target genes showed different communities of bacteria, fungi and AMF associated with the scion x rootstock combinations. Finally, all the combinations were naturally mycorrhized. The root mycorrhization intensity was influenced by the rootstock genotype, but not by the scion one. Altogether, these results suggest that both rootstock and scion genotypes influence the rhizosphere and root endophytic microbiomes. It would be interesting to analyze the biochemical composition of the rhizodeposition of these genotypes for a better understanding of the processes involved in the modulation of these microbiomes. Moreover, crossing our data with the plant agronomic characteristics could provide insights into their roles on plant fitness.

Adaptation to soil and climate through the choice of plant material

Choosing the rootstock, the scion variety and the training system best suited to the local soil and climate are the key elements for an economically sustainable production of wine. The choice of the rootstock/scion variety best adapted to the characteristics of the soil is essential but, by changing climatic conditions, ongoing climate change disrupts the fine-tuned local equilibrium. Higher temperatures induce shifts in developmental stages, with on the one hand increasing fears of spring frost damages and, on the other hand, ripening during the warmest periods in summer. Expected higher water demand and longer and more frequent drought events are also major concerns. The genetic control of the phenotypes, by genomic information but also by the epigenetic control of gene expression, offers a lot of opportunities for adapting the plant material to the future. For complex traits, genomic selection is also a promising method for predicting phenotypes. However, ecophysiological modelling is necessary to better anticipate the phenotypes in unexplored climatic conditions Genetic approaches applied on parameters of ecophysiological models rather than raw observed data are more than ever the basis for finding, or building, the ideal varieties of the future.