Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
24973 | Journal of Biotechnology | 2008 | 8 Pages |
Abstract
Directed evolution was used to expand the substrate specificity and functionality of glycine oxidase by using a high-throughput screening assay based on the 4-aminoantipyrine peroxidase system, with a coefficient of variance below 4%. After screening the library, one mutant with the desired changes was found. The mutant was purified and characterized, showing important changes compared to the wild-type, especially towards cyclic d-amino acids. Amino acid substitution of Ile15 for Val, where the consensus sequence for flavin binding site is placed, seems to be responsible for these changes in specific activity and substrate specificity. The effect of this mutation was explained by using a computer-based three-dimensional model.
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Authors
Irene Martínez-Martínez, José Navarro-Fernández, Francisco García-Carmona, Álvaro Sánchez-Ferrer,