Thermostabilization of Bacterial Fructosyl-Amino Acid Oxidase by Directed Evolution
AUTOR(ES)
Sakaue, Ryoichi
FONTE
American Society for Microbiology
RESUMO
We succeeded in isolating several thermostable mutant fructosyl-amino acid oxidase (FAOX; EC 1.5.3) without reduction of productivity by directed evolution that combined an in vivo mutagenesis and membrane assay screening system. Five amino acid substitutions (T60A, A188G, M244L, N257S, and L261M) occurred in the most thermostable mutant obtained by a fourth round of directed evolution. This altered enzyme, FAOX-TE, was stable at 45°C, whereas the wild-type enzyme was not stable above 37°C. The Km values of FAOX-TE for d-fructosyl-l-valine and d-fructosyl-glycine were 1.50 and 0.58 mM, respectively, in contrast with corresponding values of 1.61 and 0.74 mM for the wild-type enzyme. This altered FAOX-TE will be useful in the diagnosis of diabetes.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=152437Documentos Relacionados
- Functional Analysis of Fructosyl-Amino Acid Oxidases of Aspergillus oryzae
- Distribution and properties of fructosyl amino acid oxidase in fungi.
- Review of Fructosyl Amino Acid Oxidase Engineering Research: A Glimpse into the Future of Hemoglobin A1c Biosensing
- Amino acid oxidase of leukocytes in relation to H2O2-mediated bacterial killing
- The l-Amino Acid Oxidase of Molds 1
