title: Analyzing the D-amino acid content in biological samples by engineered enzymes.
abstract: The aim of our present research is to produce mutant forms of D-amino acid oxidase from Rhodotorula gracilis in order to determine D-amino acid content in different biological samples. During the past few years, our group has produced yeast D-amino acid oxidase variants with altered substrate specificity (e.g., active on acidic, or hydrophobic, or on all D-amino acids) both by rational design and directed evolution methods. Now, the kinetic constants for a number of amino acids (even for unnatural ones) of the most relevant D-amino acid oxidase variants have been investigated. This information constitutes the basis for considering potential analytical applications in this important field.
authors.affiliation: Dipartimento di Biotecnologie e Scienze Molecolari, Università degli Studi dell'Insubria, Varese, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Frattini
authors.forename: Elena
authors.initials: E
authors.lastname: Rosini
authors.forename: Loredano
authors.initials: L
authors.lastname: Pollegioni
authors.forename: Mirella S
authors.initials: MS
authors.lastname: Pilone
mesh_terms: D019943:Amino Acid Substitution; D000596:Amino Acids / Q000032:analysis* / Q000378:metabolism; D015374:Biosensing Techniques / Q000295:instrumentation*; D002611:Cheese / Q000032:analysis; D003605:D-Amino-Acid Oxidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D012248:Rhodotorula / Q000201:enzymology / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D000596:Amino Acids; D003605:D-Amino-Acid Oxidase