Development of a high-throughput screening method for racemase activity and its application to the identification of alanine racemase variants with activity towards L-arginine

作者：Willies Simon C; White Jemma L; Turner Nicholas J^*

来源：Tetrahedron, 2012, 68(37): 7564-7567.

DOI：10.1016/j.tet.2012.06.062

• Racemase
• High-throughput screening
• Protein engineering
• Biocatalysis
• Amino acid

A high-throughput solid phase screening method has been developed for alanine racemase. Conversion of an L-amino acid to the D-amino acid is detected by a D-amino acid oxidase, which is utilised as a reporter enzyme along with horseradish peroxidase and a colourimetric dye. This system has successfully been used to identify a mutant with increased activity towards arginine. Kinetic analysis and modelling studies have been performed to understand the effect of the mutations found.

• 出版日期2012-9-16