Quantification of chitinase and thaumatin-like proteins in grape juices and wines.
Identifieur interne : 000260 ( Main/Curation ); précédent : 000259; suivant : 000261Quantification of chitinase and thaumatin-like proteins in grape juices and wines.
Auteurs : D. Le Bourse [France] ; A. Conreux ; S. Villaume ; P. Lameiras ; J-M Nuzillard ; P. JeandetSource :
- Analytical and bioanalytical chemistry [ 1618-2650 ] ; 2011.
Descripteurs français
- KwdFr :
- MESH :
- analyse : Boissons, Vin.
- composition chimique : Vitis.
- isolement et purification : Chitinase, Protéines végétales.
- Chromatographie en phase liquide à haute performance, Résonance magnétique nucléaire biomoléculaire, Test ELISA.
English descriptors
- KwdEn :
- MESH :
- chemical , isolation & purification : Chitinases, Plant Proteins.
- analysis : Beverages, Wine.
- chemistry : Vitis.
- Chromatography, High Pressure Liquid, Enzyme-Linked Immunosorbent Assay, Nuclear Magnetic Resonance, Biomolecular.
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.
DOI: 10.1007/s00216-011-4912-8
PubMed: 21465097
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pubmed:21465097Le document en format XML
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<term>Chitinases (isolation & purification)</term>
<term>Chromatography, High Pressure Liquid (MeSH)</term>
<term>Enzyme-Linked Immunosorbent Assay (MeSH)</term>
<term>Nuclear Magnetic Resonance, Biomolecular (MeSH)</term>
<term>Plant Proteins (isolation & purification)</term>
<term>Vitis (chemistry)</term>
<term>Wine (analysis)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Boissons (analyse)</term>
<term>Chitinase (isolement et purification)</term>
<term>Chromatographie en phase liquide à haute performance (MeSH)</term>
<term>Protéines végétales (isolement et purification)</term>
<term>Résonance magnétique nucléaire biomoléculaire (MeSH)</term>
<term>Test ELISA (MeSH)</term>
<term>Vin (analyse)</term>
<term>Vitis (composition chimique)</term>
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<keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Chitinases</term>
<term>Plant Proteins</term>
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<keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Boissons</term>
<term>Vin</term>
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<keywords scheme="MESH" qualifier="analysis" xml:lang="en"><term>Beverages</term>
<term>Wine</term>
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<keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Vitis</term>
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<keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Vitis</term>
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<term>Protéines végétales</term>
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<keywords scheme="MESH" xml:lang="en"><term>Chromatography, High Pressure Liquid</term>
<term>Enzyme-Linked Immunosorbent Assay</term>
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<keywords scheme="MESH" xml:lang="fr"><term>Chromatographie en phase liquide à haute performance</term>
<term>Résonance magnétique nucléaire biomoléculaire</term>
<term>Test ELISA</term>
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<front><div type="abstract" xml:lang="en">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.</div>
</front>
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<Abstract><AbstractText>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.</AbstractText>
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