Substrate specificity of Amadoriase I from Aspergillus fumigatus. (Articolo in rivista)

Type
Label
  • Substrate specificity of Amadoriase I from Aspergillus fumigatus. (Articolo in rivista) (literal)
Anno
  • 2005-01-01T00:00:00+01:00 (literal)
Alternative label
  • Mennella C., Borrelli R.C., Vinale F., Ruocco M., Fogliano V. Mennella C., Borrelli R.C., Vinale F., Ruocco M., and Fogliano V. (2005)
    Substrate specificity of Amadoriase I from Aspergillus fumigatus.
    in Annals of the New York Academy of Sciences
    (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
  • Mennella C., Borrelli R.C., Vinale F., Ruocco M., Fogliano V. Mennella C., Borrelli R.C., Vinale F., Ruocco M., and Fogliano V. (literal)
Pagina inizio
  • 837 (literal)
Pagina fine
  • 844 (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume
  • 1043 (literal)
Rivista
Note
  • ISI Web of Science (WOS) (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
  • -Ruocco M., Vinale F., Istituto per la Protezione delle Piante, CNR, Sez. di Portici, Via Università 100, I-80055 Portici (NA), Italy -Mennella C., Borrelli R.C., Fogliano V., Dipartimento di Scienza degli Alimenti, Università di Napoli \"Federico II,\" via Università 100, 80055 Portici, Italy - Ruocco M., CNR, Ist. Protezione Piante - Sez. di Portici (NA) (literal)
Titolo
  • Substrate specificity of Amadoriase I from Aspergillus fumigatus. (literal)
Abstract
  • Generation of Amadori products is the major single modification by the Maillard reaction in vivo and a source of biologically active glycoxidation products leading to protein cross-linking in biological tissues. Amadoriase I from Aspergillus fumigatus cleaves Amadori products into deoxyglucosone, hydrogen peroxide, and the corresponding primary amine. It has been reported that Amadori products formed on free amino acids are a good substrate for amadoriase I, whereas the enzyme is unable to cleave Amadori products formed on whole proteins. This work aims to investigate the affinity of amadoriase I for oligopeptides and small proteins. Recombinant amadoriase I was expressed in E. coli and purified by Ni His-tag affinity chromatography. Di-, tri-, and tetrapeptides were derivatized with glucose, and the corresponding Amadori products were purified by TLC and HPLC. Glycated beta-lactoglobulin was also used as a substrate. In both cases the formation of Amadori products was monitored by electrospray ionization-mass spectrometry (ESI-MS). The Km of amadoriase for glycated-L-lysine was 4.2 mM, which is in accordance with the literature. Km decreases with the length of peptide, being slightly reduced for dipeptides, and is around 10 mM for tri- and tetrapeptides. Glycated proteins are not substrates of the enzyme; but when amadoriase I was added during the glycation reaction, a significant reduction of Amadori product formation was observed on both peptides and proteins. Data confirm the hypothesis that steric hindrance is critical for amadoriase I activity, indicating that oligopeptides up to four amino acids in length are good substrates. Moreover, these data show that, at least in some experimental conditions, amadoriase I is able to reduce the formation of protein-bound Amadori product. (literal)
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