Bacterial Expression and HTS Assessment of Soluble Epoxide Hydrolase Phosphatase

J Biomol Screen. 2016 Aug;21(7):689-94. doi: 10.1177/1087057116637609. Epub 2016 Mar 23.


Soluble epoxide hydrolase (sEH) is a bifunctional enzyme that possesses an epoxide hydrolase and lipid phosphatase activity (sEH-P) at two distinct catalytic domains. While the physiological role of the epoxide hydrolase domain is well understood, the consequences of the phosphatase activity remain unclear. Herein we describe the bacterial expression of the recombinant N-terminal domain of sEH-P and the development of a high-throughput screening protocol using a sensitive and commercially available substrate fluorescein diphosphate. The usability of the assay system was demonstrated and novel inhibitors of sEH-P were identified.

Keywords: enzyme assay; high-throughput screening; phosphatase; soluble epoxide hydrolase.

MeSH terms

  • Animals
  • Catalytic Domain / genetics
  • Enzyme Inhibitors / isolation & purification*
  • Enzyme Inhibitors / pharmacology
  • Epoxide Hydrolases / antagonists & inhibitors*
  • Escherichia coli / genetics
  • Gene Expression Regulation, Enzymologic / genetics
  • High-Throughput Screening Assays / methods*
  • Humans
  • Mice
  • Phosphoric Monoester Hydrolases / antagonists & inhibitors*
  • Phosphoric Monoester Hydrolases / genetics
  • Solubility


  • Enzyme Inhibitors
  • Phosphoric Monoester Hydrolases
  • Epoxide Hydrolases
  • EPHX2 protein, human