Journal article


Isolation, Purification and Characterization of Amadoriase Isoenzymes (Fructosyl Amine: Oxygen Oxidoreductase EC 1.5.3.) from Aspergillus sp


Publication Details
Author(s): Takahashi M, Pischetsrieder M, Monnier V
Publisher: American Society for Biochemistry and Molecular Biology
Publication year: 1997
Volume: 272
Pages range: 3437-3443
ISSN: 0021-9258

Abstract

Four 'amadoriase' enzyme fractions, which oxidatively degrade glycated low molecular weight amines and amino acids under formation of hydrogen peroxide and glucosone, were isolated from an Aspergillus sp. soil strain selected on fructosyl adamantanamine as sole carbon source. The enzymes were purified to homogeneity using a combination of ion exchange, hydroxyapatite, gel filtration, and Mono Q column chromatography. Molecular masses of amadoriase enzymes Ia, Ib, and Ic were 51 kDa, and 49 kDa for amadoriase II. Apparent kinetic constants for N(ε)-fructosyl N(α)-t-butoxycarbonyl lysine and fructosyl adamantanamine were almost identical for enzymes Ia, Ib, and Ic, but corresponding values for enzyme II were significantly different. FAD was identified in all enzymes based on its typical absorption spectrum. N- terminal sequence was identical for enzymes Ia and Ib (Ala-Pro-Ser-Ile-Leu- Ser-Thr-Glu-Ser-Ser-Ile-Ile-Val-Ile-Gly-Ala-Gly-Thr-Trp-Gly-) and Ic except that the first 5 amino acids were truncated. The sequence of enzyme II was different (Ala-Val-Thr-Lys-Ser-Ser-Ser-Leu-Leu-Ile-Val-Gly-Ala-Gly-Thr-Trp- Gly-Thr-Ser-Thr-). All enzymes had the FAD cofactor-binding consensus sequence Gly-X-Gly-X-X-Gly within the N-terminal sequence. In summary, these data show the presence of two distinct amadoriase enzymes in the Aspergillus sp. soil strain selected on fructosyl adamantanamine and induced by fructosyl propylamine. In contrast to previous described enzymes, these novel amadoriase enzymes can deglycate both glycated amines and amino acids.



How to cite
APA: Takahashi, M., Pischetsrieder, M., & Monnier, V. (1997). Isolation, Purification and Characterization of Amadoriase Isoenzymes (Fructosyl Amine: Oxygen Oxidoreductase EC 1.5.3.) from Aspergillus sp. Journal of Biological Chemistry, 272, 3437-3443. https://dx.doi.org/10.1074/jbc.272.6.3437

MLA: Takahashi, Motoko, Monika Pischetsrieder, and Vincent Monnier. "Isolation, Purification and Characterization of Amadoriase Isoenzymes (Fructosyl Amine: Oxygen Oxidoreductase EC 1.5.3.) from Aspergillus sp." Journal of Biological Chemistry 272 (1997): 3437-3443.

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