The bioactive lipid sphingosine-1-phosphate (S1P) has emerged as key regulator in cancer progression by modulating a variety of cellular processes, such as proliferation, migration, platelet aggregation, or angiogenesis. Sphingosine 1- phosphate lyase (SGPL1) irreversibly cleaves sphingosine 1-phosphate (S1P) into hexadecenal and ethanolamine phosphate, thereby controlling the concentrations of S1P. In mammalian cells, SGPL1 also targets dihydroS1P (dhS1P), a saturated form of S1P, whose precursor dihydrosphigonise is exclusively produced through de novo synthesis pathway from serine and palmitoyl condensation. These two lipids are being used interchangeably in the assay for SGPL1 enzymatic activity. However, dhS1P and S1P exhibit opposite effects in certain cellular processes and their levels respond differently to environmental factors including cancer therapies. In order to better understand the enzymatic kinetics of SGPL1 toward these two substrates, we cloned a coding sequence of human SGPL1 (hSGPL1) into a bacterial expression system pQ60 vector. In further studies, this material will provide the material for the expression and purification of recombinant human SGPL1 enzyme, which will then be used to study its specificity and efficacy on dhS1P and S1P
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