Studies on the effect of amadoriase from Aspergillus fumigatus on peptide and protein glycation in vitro

Edoardo Capuano, Francalisa Fedele, Carmela Mennella, Marianna Visciano, Aurora Napolitano, Stefania Lanzuise, Michelina Ruocco, Matteo Lorito, María Dolores Del Castillo, Vincenzo Fogliano*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

10 Citations (Scopus)

Abstract

Amadoriase I is a fructosyl amine oxidase from Aspergillus fumigatus that catalyzes the oxidation of Amadori products (APs) producing glucosone, H 2O2, and the corresponding free amine. All the enzymes of this family discovered so far only deglycate small molecular weight products and are inactive toward large molecular weight substrates, such as glycated BSA or ribonuclease A. Therefore, they cannot be used to reverse protein glycation occurring in diabetes or in foods. In this paper, the effect of Amadoriase I added during the in vitro reaction between glucose and peptides having different polarities or proteins with molecular weights ranging from to 5 to 66 kDa was tested. The formation of APs was monitored by ESI-MS of intact glycated protein or peptides and by measuring the Nε-(1-deoxy-D-fructos-1-yl)-L- lysine and furosine concentrations. Results showed that the formation of APs is reduced up to 80% when peptides and glucose are incubated in the presence of Amadoriase. The effect is more evident for hydrophobic peptides. In protein-glucose systems, the effect was dependent on the molecular weight and steric hindrance being negligible for BSA and at a maximum for insulin, where the formation of APs was reduced up to 60%. These findings indicate new potential applications of Amadoriase I as an efficient tool for inhibiting protein glycation in real food systems.

Original languageEnglish
Pages (from-to)4189-4195
Number of pages7
JournalJournal of Agricultural and Food Chemistry
Volume55
Issue number10
DOIs
Publication statusPublished - 16 May 2007
Externally publishedYes

Keywords

  • Amadori products
  • Amadoriase I
  • Browning
  • Protein glycation

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