title: Distribution and properties of novel deglycating enzymes for fructosyl peptide in fungi.
abstract: Our fungal culture collection was screened for fructosyl peptide oxidase, an enzyme that could be used for the determination of glycated hemoglobin in diabetic subjects with hyperglycemia. Fructosyl peptide oxidases were found in strains of eight genera: Achaetomiella, Achaetomium, Chaetomium, Coniochaeta, Eupenicillium, Gelasinospora, Microascus and Thielavia. By their substrate specificity toward N(alpha)-fructosyl valyl-histidine (alpha-keto-amine) and N(epsilon)-fructosyl lysine (epsilon-keto-amine), fructosyl peptide oxidases could be categorized into two groups: (1) enzymes that oxidize both alpha-keto-amine and epsilon-keto-amine, and (2) enzymes that preferably oxidize alpha-keto-amine. A fructosyl peptide oxidase from Achaetomiella virescens ATCC 32393, active toward both N(alpha)-fructosyl valyl-histidine and N(epsilon)-fructosyl lysine, was purified to homogeneity and characterized. The enzyme was monomeric ( M(r)=50,000), was most active at 40 degrees C and pH 8.0, and had a covalently bound flavin as a prosthetic group. Apparent K(m) values for N(alpha)-fructosyl valyl-histidine and N(epsilon)-fructosyl lysine were 2.30 and 1.69 mM, respectively. N(alpha)-fructosyl valyl-histidine was consumed and the same molar amount of valyl-histidine was produced by the fructosyl peptide oxidase reaction. This enzyme could be useful for the measurement of hemoglobin A(1C), the N-terminal valine residue of the beta-subunit of which is glycated.
authors.affiliation: Research and Development Division, Kikkoman Corporation, 399 Noda, Chiba Pref. 278-0037, Noda-city, Japan. khirokawa@mail.kikkoman.co.jp
authors.forename: Kozo
authors.initials: K
authors.lastname: Hirokawa
authors.forename: Keiko
authors.initials: K
authors.lastname: Gomi
authors.forename: Mikio
authors.initials: M
authors.lastname: Bakke
authors.forename: Naoki
authors.initials: N
authors.lastname: Kajiyama
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D006442:Glycated Hemoglobin / Q000031:analogs & derivatives* / Q000378:metabolism; D006639:Histidine / Q000031:analogs & derivatives / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008239:Lysine / Q000031:analogs & derivatives* / Q000378:metabolism; D020037:Sordariales / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature; D014633:Valine / Q000031:analogs & derivatives* / Q000378:metabolism
chemical_list: D000594:Amino Acid Oxidoreductases; D006442:Glycated Hemoglobin; D006639:Histidine; D008239:Lysine; D014633:Valine; C025617:glucosylated hemoglobin A; C045420:fructosylvaline; C033186:fructosyl-lysine; C520096:fructosyl-peptide oxidase