Continuous High-Throughput Colorimetric Assays for α-Transaminases

Methods Mol Biol. 2018:1685:233-245. doi: 10.1007/978-1-4939-7366-8_13.

Abstract

Transaminases are efficient tools for the stereoselective conversion of prochiral ketones into valuable chiral amines. Notably, the diversity of naturally occurring α-transaminases offers access to a wide range of L- and D-α-amino acids. We describe here two continuous colorimetric assays for the quantification of transamination activities between a keto acid and a standard donor substrate (L- or D-Glutamic acid or cysteine sulfinic acid). These assays are helpful for kinetic studies as well as for high-throughput screening of enzyme collections.

Keywords: Amino acids; Cysteine sulfinic acid; High-throughput screening; Screening assay; Transaminase.

MeSH terms

  • Colorimetry
  • Cysteine / analogs & derivatives
  • Cysteine / metabolism
  • Glutamic Acid / metabolism
  • High-Throughput Screening Assays / methods*
  • Keto Acids / metabolism
  • Transaminases / metabolism*

Substances

  • Keto Acids
  • Glutamic Acid
  • Transaminases
  • Cysteine
  • cysteine sulfinic acid