Release of protein-bound adducts of cysteine residues with caffeic acid by a modified enzymatic hydrolysis method using Pronase E

Research output: Contribution to journal › Article › Research › peer review

Authors

  • Raphaela Krax
  • Leon Schulte
  • Annik Fischer
  • Michael Hellwig

External Research Organisations

  • Technische Universität Braunschweig
  • Technische Universität Dresden
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Details

Original languageEnglish
Article number143379
JournalFood chemistry
Volume476
Early online date14 Feb 2025
Publication statusE-pub ahead of print - 14 Feb 2025
Externally publishedYes

Abstract

Polyphenols can be widely found in plants and plant-based food. Polyphenols with an ortho-dihydroxy structure, such as caffeic acid (CA), can easily be oxidized to their ortho-quinones. These quinones can react with nucleophilic side chains of proteins, such as the thiol group in cysteine (Cys), and form adducts that can affect the structure and function of these proteins. Individual adducts have been described qualitatively, but quantitative analysis has been hampered by the poor stability of the adducts during sample preparation. In this study, 2-S-cysteinyl caffeic acid was synthesized either in a free or protein-bound form. A one-enzyme hydrolysis was developed using Pronase E. It yielded a hydrolysis rate of 97 % calculated via phenylalanine release. A recovery rate of 2-S-cysteinyl caffeic acid (2-CCA) of 64 % after hydrolysis was achieved when buffer systems containing borate anions were used. The formation of 2-CCA in bovine serum albumin (BSA) incubated with CA was shown.

Keywords

    2-S-Cysteinyl caffeic acid, Bovine serum albumin, Pronase E, Protein hydrolysis, Protein-polyphenol interaction

ASJC Scopus subject areas

Cite this

Release of protein-bound adducts of cysteine residues with caffeic acid by a modified enzymatic hydrolysis method using Pronase E. / Krax, Raphaela; Schulte, Leon; Fischer, Annik et al.
In: Food chemistry, Vol. 476, 143379, 01.06.2025.

Research output: Contribution to journal › Article › Research › peer review

Krax R, Schulte L, Fischer A, Hellwig M. Release of protein-bound adducts of cysteine residues with caffeic acid by a modified enzymatic hydrolysis method using Pronase E. Food chemistry. 2025 Jun 1;476:143379. Epub 2025 Feb 14. doi: 10.1016/j.foodchem.2025.143379
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AU - Krax, Raphaela

AU - Schulte, Leon

AU - Fischer, Annik

AU - Hellwig, Michael

N1 - Publisher Copyright: © 2025 The Authors

PY - 2025/2/14

Y1 - 2025/2/14

N2 - Polyphenols can be widely found in plants and plant-based food. Polyphenols with an ortho-dihydroxy structure, such as caffeic acid (CA), can easily be oxidized to their ortho-quinones. These quinones can react with nucleophilic side chains of proteins, such as the thiol group in cysteine (Cys), and form adducts that can affect the structure and function of these proteins. Individual adducts have been described qualitatively, but quantitative analysis has been hampered by the poor stability of the adducts during sample preparation. In this study, 2-S-cysteinyl caffeic acid was synthesized either in a free or protein-bound form. A one-enzyme hydrolysis was developed using Pronase E. It yielded a hydrolysis rate of 97 % calculated via phenylalanine release. A recovery rate of 2-S-cysteinyl caffeic acid (2-CCA) of 64 % after hydrolysis was achieved when buffer systems containing borate anions were used. The formation of 2-CCA in bovine serum albumin (BSA) incubated with CA was shown.

AB - Polyphenols can be widely found in plants and plant-based food. Polyphenols with an ortho-dihydroxy structure, such as caffeic acid (CA), can easily be oxidized to their ortho-quinones. These quinones can react with nucleophilic side chains of proteins, such as the thiol group in cysteine (Cys), and form adducts that can affect the structure and function of these proteins. Individual adducts have been described qualitatively, but quantitative analysis has been hampered by the poor stability of the adducts during sample preparation. In this study, 2-S-cysteinyl caffeic acid was synthesized either in a free or protein-bound form. A one-enzyme hydrolysis was developed using Pronase E. It yielded a hydrolysis rate of 97 % calculated via phenylalanine release. A recovery rate of 2-S-cysteinyl caffeic acid (2-CCA) of 64 % after hydrolysis was achieved when buffer systems containing borate anions were used. The formation of 2-CCA in bovine serum albumin (BSA) incubated with CA was shown.

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