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Quantification of chitinase and thaumatin-like proteins in grape juices and wines

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An Erratum to this article was published on 08 May 2011

Abstract

Chitinases and thaumatin-like proteins are important grape proteins as they have a great influence on wine quality. The quantification of these proteins in grape juices and wines, along with their purification, is therefore crucial to study their intrinsic characteristics and the exact role they play in wines. The main isoforms of these two proteins from Chardonnay grape juice were thus purified by liquid chromatography. Two fast protein liquid chromatography (FLPC) steps allowed the fractionation and purification of the juice proteins, using cation exchange and hydrophobic interaction media. A further high-performance liquid chromatography (HPLC) step was used to achieve higher purity levels. Fraction assessment was achieved by mass spectrometry. Fraction purity was determined by HPLC to detect the presence of protein contaminants, and by nuclear magnetic resonance (NMR) spectroscopy to detect the presence of organic contaminants. Once pure fractions of lyophilized chitinase and thaumatin-like protein were obtained, ultra-HPLC (UHPLC) and enzyme-linked immunosorbent assay (ELISA) calibration curves were constructed. The quantification of these proteins in different grape juice and wine samples was thus achieved for the first time with both techniques through comparison with the purified protein calibration curve. UHPLC and ELISA showed very consistent results (less than 16% deviation for both proteins) and either could be considered to provide an accurate and reliable quantification of proteins in the oenology field.

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Abbreviations

BSA:

bovine serum albumin

Cyt C:

horse heart cytochrome C

FPLC:

fast protein liquid chromatography

HIC:

hydrophobic interaction

HPLC:

high-performance liquid chromatography

MES:

2-(N-morpholino)ethane sulfonic acid

NMR:

nuclear magnetic resonance

PBS:

phosphate-buffered saline

PR:

pathogenesis-related

PTA-ELISA:

plate-trapping antigen–enzyme-linked immunosorbent assay

RP:

reversed-phase

RSD:

relative standard deviation

SCX:

strong cation exchange

SDS-PAGE:

sodium dodecyl sulfate–polyacrylamide gel electrophoresis

TBS:

Tris-buffered saline

TFA:

trifluoroacetic acid

TL:

thaumatin-like

UHPLC:

ultra-HPLC

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Acknowledgements

We thank the Champagne Ardenne Area, the General Council of Marne, the Ministry of Research, the Ville de Reims, the A.R.O.C.U. Association, and the Champagne Moët & Chandon for their support. We gratefully thank the wine cooperative of Nogent l’Abesse (France) for providing grape juice and wines samples. We also acknowledge Dr. S. Jégou (University of Reims Champagne-Ardenne, France) in appreciation of her contribution.

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Authors and Affiliations

  1. Laboratoire d’Oenologie et Chimie Appliquée, Université de Reims Champagne-Ardenne, URVVC-SE UPRES EA 2069, BP 1039, 51687, Reims Cedex 2, France

    D. Le Bourse, A. Conreux, S. Villaume & P. Jeandet

  2. CNRS, Institut de Chimie Moléculaire de Reims, BP 1039, 51687, Reims Cedex 2, France

    P. Lameiras & J.-M. Nuzillard

Authors
  1. D. Le Bourse
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  2. A. Conreux
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  3. S. Villaume
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  4. P. Lameiras
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  5. J.-M. Nuzillard
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  6. P. Jeandet
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Corresponding author

Correspondence to D. Le Bourse.

Additional information

Published in the special issue Wine Analysis with Guest Editor Isabelle Pianet.

An erratum to this article can be found at http://dx.doi.org/10.1007/s00216-011-5053-9

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Le Bourse, D., Conreux, A., Villaume, S. et al. Quantification of chitinase and thaumatin-like proteins in grape juices and wines. Anal Bioanal Chem 401, 1541–1549 (2011). https://doi.org/10.1007/s00216-011-4912-8

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  • DOI: https://doi.org/10.1007/s00216-011-4912-8

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