Analyzing the D-amino acid content in biological samples by engineered enzymes

作者：Frattini Luca; Rosini Elena; Pollegioni Loredano^*; Pilone Mirella S

来源：Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2011, 879(29): 3235-3239.

DOI：10.1016/j.jchromb.2011.02.036

• Substrate specificity
• D-Amino acids
• Detection
• Flavoproteins
• Oxidases
• Mutagenesis

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.

• 出版日期2011-11-1