M. García,
R. Apolinar-Valiente, P. Williams,
J. Crespo,
B. Esteve-Zarzoso, J. M. Cabellos,
T. Arroyo, T. Doco
18 July 2017
Influence on complex carbohydrates of Malvar white
wines elaborated with Torulaspora delbrueckii and
Saccharomyces cerevisiae native yeasts
D. O. “Vinos de Madrid”, 3 subzones:
-Arganda
-Navalcarnero
-San Martín de Valdeiglesias
IMIDRA Yeast Collection: CLI Collection
Grapes:
•White
•Originally from Madrid
•Yellowish-Green Berry
Must:
•Low pH
•Medium acidity
Wine:
•Light and fresh wine
•Medium-high alcoholic degree
•High content in volatile higher alcohols
Malvar Grape Variety
Selected native yeast strains:
•Saccharomyces cerevisiae CLI 889
•Torulaspora delbrueckii CLI 918
Cultures:
•Pure culture of S. cerevisiae (CONTROL): p-Sc
•Pure culture of T. delbrueckii: p-Td
•Mixed culture of T. delbrueckii + S. cerevisiae (simultaneously inoculated): m-Td/Sc
•Sequential culture of T. delbrueckii + S. cerevisiae (Saccharomyces inoculated with 5°
Alcohol): s-Td/Sc
Fermentations
Liberated during the pressing, the winemaking process and the aging on lees, they come from grape cell walls and microorganisms cell walls
Multiple functions with oenological importance
One of the main group of macromolecules present in musts and wines
Polysaccharides in wine have been largely studied
Several physicochemical and sensorial properties of interest
Origin: pectin and hemicellulosic structures of grapes, but also from yeasts cell walls
Oligosaccharides in wine have been recently studied
This work focuses on determination of polysaccharides and oligosaccharides release into Malvar white wines, elaborated with T. delbrueckii CLI 918 and S. cerevisiae CLI 889 native yeasts from D.O. “Vinos de Madrid”,
in pure, mixed and sequential cultures made with both
strains
High-Resolution Size-Exclusion Chromatography
Superdex- 30HR1 column
Oligosaccharide s collection
RI detector
Polysaccharides collection
Isolation of polysaccharide and oligosaccharide fractions:
Molar mass distributions and intrinsic viscosity of Polysaccharides: SEC-MALLS.
Glycosyl-linkage determination of Polysaccharides:
by GC-MS of the partially methylated alditol acetates
Analytical methods
Oligosaccharide composition:
by GC of their TMS derivatives.
Rhamnose
Arabinose
Galactose
Glucose
Mannose
Mole percentage
10 0 20 30 40 50 60 p-Sc
Rhamnose
Arabinose
Galactose
Glucose
Mannose
Mole percentage
10 0 20 30 40 50 60
Rhamnose
Arabinose
Galactose
Glucose
Mannose
Mole percentage
10 0 20 30 40 50 60 m-Td/Sc
*
Rhamnose
Arabinose
Galactose
Glucose
Mannose
Mole percentage
10 0 20 30 40 50 60 *
*
s-Td/Sc
*
*
p-Td *
Glycosyl-Linkage Composition of Polysaccharides Fraction
*
*
*
* * * *
*
*
*
Characteristic of branched arabinans on the RG chains of pectins
Ara linked at positions 2 and 3 arising from AGPs from grapes.
Gal linked at 3, 6,
3,6and3,4,6 linkages are found in AGPs.
Glycosyl-Linkage Composition of Polysaccharides Fraction
Ratio 3- / 3,6-linked Gal represented a highly- branched AGP molecule in Malvar wines
*
*
* *
*
*
*
*
Ratio Terminal/2-linked Man in p-Td has value of 4 thus MPs liberated by Td strain contain 4 residues of Man linked in 2 for 1 Terminal Man.
MPs structure similar in p-Td and s-Td/Sc MPs structure similar in
p-Sc and m-Td/Sc
Glycosyl-Linkage Composition of Polysaccharides Fraction
Peak 1
Peak 2
Peak 3
M o la r m as s (g /m o l) R e la ti ve s ca le
Time (minutes)
SEC-MALLS Analysis of Polysaccharides Fraction
Molar mass (Mw) was higher by p-Td in all three populations
First populations correspond to MPs (Vidal et al., 2003)
SEC-MALLS Analysis of Polysaccharides Fraction
Intrinsic viscosity was notably lower for p-Td and s-Td/Sc
SEC-MALLS Analysis of Polysaccharides Fraction
SEC-MALLS Analysis of Polysaccharides Fraction
Range 1
•0 % in p-Td
•In all Malvar wines, values below the described in red wines
•Corresponds to MPs of small molar mass and mainly RG-II in red wines
SEC-MALLS Analysis of Polysaccharides Fraction
Range 2
•p-Td > m-Td/Sc > p-Sc > s-Td/Sc
•Corresponds to MPs of small molar mass
SEC-MALLS Analysis of Polysaccharides Fraction
Range 3
•s-Td/Sc > p-Td > m-Td/Sc > pSc
•Corresponds to MPs of medium molar mass and AGPs
SEC-MALLS Analysis of Polysaccharides Fraction
SEC-MALLS Analysis of Polysaccharides Fraction
Range 4, 5 and 6
•Similar values between four different types of cultures
Rh am no se Ar ab in os e
Ga la ct os e Xy lo se
Ga l A Gl c A 4- O- M eG lc A
Fu co se
Xy lit ol Glc M an
M o le p e rc e n ta ge
10 0 20 30 40 50
Oligosaccharide Fraction
Rh am no se Ar ab in os e
Ga la ct os e Xy lo se
Ga l A Gl c A 4- O- M eG lc A
Fu co se
Xy lit ol Glc M an
M o le p e rc e n ta ge
10 0 20 30 40 50
Oligosaccharide Fraction
Rh am no se Ar ab in os e
Ga la ct os e Xy lo se
Ga l A Gl c A 4- O- M eG lc A
Fu co se
Xy lit ol Glc M an
M o le p e rc e n ta ge
10 0 20 30 40 50
Oligosaccharide Fraction
Rh am no se Ar ab in os e
Ga la ct os e Xy lo se
Ga l A Gl c A 4- O- M eG lc A
Fu co se
Xy lit ol Glc M an
M o le p e rc e n ta ge
10 0 20 30 40 50
Oligosaccharide Fraction
Rh am no se Ar ab in os e
Ga la ct os e Xy lo se
Ga l A Gl c A 4- O- M eG lc A
Fu co se
Xy lit ol Glc M an
M o le p e rc e n ta ge
10 0 20 30 40 50
Oligosaccharide Fraction
Ara/Gal Ratio
•Characteristic of wine PRAGs
•Higher values in p-Sc and m-Td/Sc
•In red wines, values described close to 1
Oligosaccharide Fraction
Rha/Gal A Ratio
•Relative richness of the wine oligosaccharides in HGs vs RGs
•Low values in Malvar wines
•Values lower than those found in red still wines
Oligosaccharide Fraction
(Ara+Gal)/Rha Ratio
•Relative importance of the neutral side chains to the RG backbone
•Higher ratio in p-Sc culture
•Cultures elaborated with T. delbrueckii contain more structures from the hairy regions of pectins (RG-like structures) due to breakdown of grape cell wall berries.
Oligosaccharide Fraction
Man/Glc Ratio
•Effectiveness of MPs for protein stabilization in white wines
•In all cultures, Glc was the major residue sugar arising from cellulose and hemicellulose
xyloglucans of grape berries (Vidal et al., 2001) or from yeast glucans released during alcoholic fermentation.
