Purification and Properties of Fructosylamine Oxidase from<i>Aspergillus</i>sp. 1005

Horiuchi, Tatsuo; Kurokawa, Toshiko

doi:10.1080/00021369.1991.10870584

Cited by 17 publications

(19 citation statements)

References 3 publications

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“…The first fully characterized enzyme was described by Horiuchi and colleagues (31) in Corynebacterium sp., followed by a similar enzyme from Aspergillus sp. by the same group (32). Both enzymes regenerate the free amino acid and produce H 2 O 2 and glucosone.…”

Section: Search For An Amadori Product Degrading Microorganismmentioning

confidence: 99%

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

Takahashi¹,

Pischetsrieder²,

Monnier

1997

Journal of Biological Chemistry

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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 ⑀ -fructosyl N ␣ -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-SerThr-Glu-Ser-Ser-Ile-Ile-Val-Ile-Gly-Ala-Gly-Thr-TrpGly-) and Ic except that the first 5 amino acids were truncated. The sequence of enzyme II was different (AlaVal-Thr-Lys-Ser-Ser-Ser-Leu-Leu-Ile-Val-Gly-Ala-GlyThr-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.

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Section: Search For An Amadori Product Degrading Microorganismmentioning

confidence: 99%

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

Takahashi¹,

Pischetsrieder²,

Monnier

1997

Journal of Biological Chemistry

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“…[16] and Aspergillus sp. [17]. The enzymes showed low activities toward fructosyl lysine and bound to FAD non-covalently.…”

Section: Discussionmentioning

confidence: 99%

Primary Structures of Fungal Fructosyl Amino Acid Oxidases and their Application to the Measurement of Glycated Proteins

Yoshida

Sakai

Isogai

et al. 1996

European Journal of Biochemistry

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Fructosyl amino acid oxidase (FAOD), which is active toward model compounds of the glycated proteins in blood, N'-fructosyl Nu-Z-lysine and N-fructosyl valine, was purified to homogeneity from Aspergillus terreus GP1. Though the enzyme did not use glycated proteins directly as its substrate, it used glycated human serum albumin (HSA) when HSA was treated with a protease. Linear relationships between both the concentration and the increase in absorbance and the glycation rate of glycated HSA and the increase in absorbance were observed. cDNAs coding for FAODs were cloned from cDNA libraries of A. terreus GPI and Penicillium janthinellum AKU 3413. The coding region for both fungal FAODs consisted of 1314 bp encoding 437 amino acids. The sequence of a dinucleotide-binding motif, GXGXXG, was in the deduced N-terminal region and a similar sequence to that the active site of bacterial sarcosine oxidases was found near the C-terminal region of FAOD. The of C-terminal tripeptides SKL and AKL of FAODs from A. terreus and R janthinellum, respectively, represent typical peroxisomaltargeting signals. Finally, FAOD protein was produced in Escherichia coli transformants in an active form, and at the same level a? in the original fungi.Keywords: fructosyl amino acid oxidase; Amadori rearrangement; glycated protein ; Aspergillus terreus GPI ; Penicilliurn ,janthinelhm AKU3413.Reducing sugars such as glucose attach to the amino groups of proteins by forming Schiff's bases. Subsequently, the adduct undergoes Amadori rearrangement to form a stable ketoamine product [l -41. This nonenzymic reaction is called glycation, in order to distinguish it from the enzymic glycosylation of proteins. In diabetic patients whose blood glucose levels are high, the glycation of blood proteins, e.g. hemoglobin and albumin, is enhanced. The amounts of these glycated proteins reflect the level of blood glucose in periods corresponding to the half-life of the protein (14-20 days for albumin and 1-3 months for hemoglobin). Since the glycation of blood proteins is not affected by transient increases in blood glucose, the levels of the glycated proteins are good indices for monitoring diabetes mellitus patients during therapy.The prevalent methods used for the determination of glycated proteins in serum are high-performance liquid chromatography [5] and the fructosamine method [6]. The specificities of these methods are relatively low and/or the assays are somewhat tedious when examining a great number of samples. To overcome these demerits, we are developing a new enzymic method which is highly specific and easy to conduct.The most commonly glycated site of albumin is the &-amino group of the lysine residue [7] ; that of hemoglobin A,, (HbA,,) Correspondence to N. Kato, Department of Agricultural Chemistry, Faculty of Agnculture, Kyoto University, Sakyo-ku, Kyoto, Japan 606-01Abbreviations. FAOD, fructosyl amino acid oxidase; Z-Lys(Fru), Wfructocyl Nu-2-lysine ; Fru-Lys(Z), Wfructosyl N'-Z-lysine; Fru-Val, fructosyl valine; Z-, benzyloxycarbonyl-; Hb...

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“…and Aspergillus sp. by Horiuchi et al (34). These FAD-containing enzymes were found to deglycate fructosyl amino acids into glucosone and the primary amine under formation of h 2 o 2 (Fig.…”

Section: Natural Defence Mechanisms Against Ages Formationmentioning

confidence: 99%