Studies on the effect of Amadoriase from Aspergillus fumigatus on peptide and protein glycation in vitro.
Amadoriase I is a fructosyl amine oxidase from Aspergillus fumigatus that catalyzes the oxidation of Amadori products (APs) producing glucosone, H2O2, 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 Nepsilon-(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.
Capuano
Edoardo
E
Dipartimento di Scienza degli Alimenti and Dipartimento Ar.Bo.Pa.Ve. sez. Patologia Vegetale, University of Napoli Federico II, via Università 100, 80055 Portici, Italy.
Fedele
Francalisa
F
Mennella
Carmela
C
Visciano
Marianna
M
Napolitano
Aurora
A
Lanzuise
Stefania
S
Ruocco
Michelina
M
Lorito
Matteo
M
del Castillo
María Dolores
MD
Fogliano
Vincenzo
V
D000594:Amino Acid Oxidoreductases / Q000378:metabolism*; D001232:Aspergillus fumigatus / Q000201:enzymology*; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D057927:Hydrophobic and Hydrophilic Interactions; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism
D006023:Glycoproteins; D010455:Peptides; D012710:Serum Albumin, Bovine; D000594:Amino Acid Oxidoreductases; C103012:amadoriase