enzymatic method for the determination of hemoglobinA1C

• Published in: Biotechnology letters Vol. 27; no. 14; pp. 963 - 968
• Main Authors: Hirokawa, Kozo, Shimoji, Kazuhiko, Kajiyama, Naoki
• Format: Journal Article
• Language: English
• Published: Dordrecht Kluwer Academic Publishers 01.07.2005; Springer; Springer Nature B.V
• Subjects: Biological and medical sciences; Biotechnology; Fundamental and applied biological sciences. Psychology; hemoglobin; methodology; Paenibacillus polymyxa; Peptides; proteinases; fructosyl amino acid oxidase; Peptides; Enzyme; amadoriase; fructosyl peptide oxidase; Oxidase; enzymatic measurement of HbA1C; Enzymatic method; Aminoacid; Hemoglobin; Oxidoreductases; hemoglobin A1C; diabetes; Enzymatic reaction
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-005-7832-x

Fructosyl peptide oxidase is a flavoenzyme that catalyzes the oxidative deglycation of N-(1-deoxyfructosyl)-Val-His, a model compound of hemoglobin (Hb)A₁C. To develop an enzymatic method for the measurement of HbA₁C, we screened for a proper protease using N-(1-deoxyfructosyl)-hexapeptide as a substrate. Several proteases, including Neutral protease from Bacillus polymyxa, were found to release N-(1-deoxyfructosyl)-Val-His efficiently, however no protease was found to release N-(1-deoxyfructosyl)-Val. Neutral protease also digested HbA₁C to release N-(1-deoxyfructosyl)-Val-His, and then the fructosyl peptide was detected using fructosyl peptide oxidase. The linear relationship was observed between the concentration of HbA₁C and the absorbancy of fructosyl peptide oxidase reaction, hence this new method is a practical means for measuring HbA₁C.