Polysaccharides from grape berry cell walls. Part I: tissue distribution and structural characterization of the pectic polysaccharides

doi:10.1016/S0144-8617(00)00285-X

Carbohydrate Polymers

Volume 45, Issue 4, August 2001, Pages 315-323

https://doi.org/10.1016/S0144-8617(00)00285-X Get rights and content

Abstract

Buffer-soluble arabinogalactan-proteins (AGPs) and pectins from grape berry skin and pulp tissues have been isolated and their structure has been partly determined. Pectic polysaccharides from the cell wall material were solubilized by treating pulp and skin cell walls with homogeneous glycosyl hydrolases. Homogalacturonans, rhamnogalacturonans I (RG-I), and rhamnogalacturonan II (RG-II) of each tissue have been fractionated by high resolution size exclusion chromatography and their relative distribution and major structural features have been determined. It has been shown that pulp tissue contains two-fold more buffer-soluble AGPs and pectins than skin tissue and we have determined that 75% of the grape berry walls originates from the skin tissue. There is three-fold more RG-I and RG-II in skin tissue than in pulp tissue and three-fold more RG-I than RG-II in the grape berry cell walls.

The results of this study have shown that the grape polysaccharide content of a wine is related to the type of tissue used for wine making and to the solubility of the grape polysaccharides and their resistance to fragmentation by grape and yeast glycanases.

Introduction

The composition and structure of grape berry primary cell walls are of interest because of their importance in wine manufacturing technology (Gerbaud et al., 1996, Vernhet et al., 1999, Vernhet et al., 1996) and in fruit ripening (Barnavon et al., 2000, Nunan et al., 1998). The alcohol-insoluble residues (AIR) prepared from grape berry pulp (pulp tissue) consists predominantly of cellulose, the hemicellulose xyloglucan, and the pectic polysaccharides homogalacturonan (HG), rhamnogalacturonan I (RG-I), and rhamnogalacturonan II (RG-II) (Nunan et al., 1997, Saulnier and Thibault, 1987). The side-chains of RG-I are composed of arabinans and type I and type II arabinogalactans (Saulnier, Brillouet & Joseleau, 1988). The glycosyl-residue compositions of pulp and grape berry skins (skin tissue) AIR are similar (Nunan et al., 1997), although polysaccharides have been reported to account for only 50% of the skin AIR (Lecas & Brillouet, 1994).

Buffer-soluble type II arabinogalactan-proteins (AGPs) are abundant in grape berry tissue and have been partially characterized (Saulnier & Brillouet, 1989). In contrast, the pectic polysaccharide RG-II accounts for <5% of the pulp (Nunan et al., 1997) and skin (Lecas & Brillouet, 1994) cell walls. Nevertheless, RG-II and AGPs are the quantitatively major grape polysaccharides present in wine (Brillouet et al., 1990, Pellerin et al., 1995, Doco and Brillouet, 1993, Pellerin et al., 1996). Moreover, their presence in wine is believed to affect haze formation (Waters, Pellerin & Brillouet, 1994) and the binding of heavy metals (Szpunar et al., 1998).

We now report the isolation and partial structural characterization of buffer-soluble AGPs and pectins from grape berry skin and pulp tissue. Buffer-insoluble pectic polysaccharides were solubilized by treating pulp and skin cell walls with homogeneous glycosyl hydrolases. The relative distribution and major structural features of HG, RG-I, and RG-II in these walls have been determined. These studies show that the amounts of pulp and skin tissue used in wine making, the polysaccharide contents of the tissues, and the stability of the polysaccharides to enzymic fragmentation during fermentation determine the grape polysaccharide contents of red and white wine.

Section snippets

Plant material

Mature grapes of the cultivar Grenache blanc were harvested at the Unité Expérimentale d'&OElig:nologie INRA-Pech Rouge (Gruissan, France). Skin, seeds, and pulp tissues from frozen berries (3 kg) were hand-separated with a scalpel. The isolated tissues were then frozen in liquid nitrogen and stored at −80°C.

Isolation of the buffer-soluble polysaccharides from grape tissues

All steps were performed at 4°C to minimize fragmentation of wall polysaccharides by endogenous enzymes. Skin and pulp tissues (600 g of each) were suspended in 1.2 l of 40 mM HEPES, pH 7, and

The buffer-soluble polysaccharides from grape berry pulp and skin tissue

In previous studies, the wall polysaccharide content of grape berries was estimated by analysis of the residue remaining after aqueous 80% ethanol treatment of the berries (Nunan et al., 1997, Saulnier et al., 1988). This procedure does not distinguish buffer-soluble polysaccharides and proteoglycans from wall-bound polymers. In our study, grape tissue was treated with 40 mM HEPES, pH 7.0, and the buffer-soluble polysaccharides partially characterized. These polysaccharides account for 30 and

Conclusions

For the first time, grape pulp and skin buffer-soluble and pectic polysaccharides have been isolated simultaneously. We have provided evidence that pulp tissue contains two-fold more buffer-soluble AGPs and pectins than skin tissue and that 75% of the grape berry walls originates from the skin tissue. Nevertheless, comparable amounts of HG, RG-I, and RG-II are present in pulp and skin cell walls. The wall-bound pectic polysaccharides are structurally similar to those previously described. Our

Acknowledgements

We thank Dr Carl Bergmann (Complex Carbohydrate Research Center, University of Georgia, Athens, USA) for providing the enzymes and Dr Markus Pauly (Novo Nordisk Kopenhagen, Denmark) for supplying the recombinant endoglucanase. The authors wish to thank also Dr J. Visser (Wageningen, The Netherlands) and Novo Nordisk for recombinant EndoPG, ExoPG and PME gifts.

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Citation Excerpt :
Although the RG-I content in grape skin cell walls is approximately-three times higher than that of RG-II, RG-I is the least abundant polysaccharide component in wine (approximately 4 % of the total polysaccharides in red wine, Table 1). This is likely due to incomplete solubilization from the cell walls and weak resistance to enzyme hydrolysis during winemaking (Vidal et al., 2001; Wang et al., 2021a). In addition, the small amounts of xylose residues from grape berry cell walls suggest that traces of hemicelluloses are solubilized (Guadalupe et al., 2012).
Wine quality is closely related to various compounds including polysaccharides, a class of crucial macromolecules that affect its chemical and physical properties by influencing the colloidal state or interacting with other compounds via non-covalent bonds. Herein, the composition and structural characteristics of the major polysaccharides identified in wine and the factors influencing their contents are briefly described. An improved understanding of the molecular mechanisms of wine polysaccharides and their practical applications on wine stability and organoleptic qualities is thoroughly discussed. The effects of polysaccharides on wine quality are significantly correlated with their structure and composition as well as wine matrix composition. Thus, to better understand the chemical complexity of polysaccharides, relevant analytical methods are systematically summarized and highlighted, which may ultimately lead to the development of novel winery guidelines.

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Polysaccharides from grape berry cell walls. Part I: tissue distribution and structural characterization of the pectic polysaccharides

Abstract

Introduction

Section snippets

Plant material

Isolation of the buffer-soluble polysaccharides from grape tissues

The buffer-soluble polysaccharides from grape berry pulp and skin tissue

Conclusions

Acknowledgements

FEBS Letters

Analytical Biochemistry

Biochimie

Carbohydrate Research

Carbohydrate Research

Plant Physiology and Biochemistry

Carbohydrate Research

Phytochemistry

Phytochemistry

Carbohydrate Research

Journal of Biological Chemistry

Carbohydrate Research

Carbohydrate Research

Carbohydrate Polymers

Carbohydrate Research

Carbohydrate Research

Carbohydrate Research

Carbohydrate Research

A simplified method for accurate determination of cell wall uronide content

Journal of Food Biochemistry