The synthesis of ATP in the human parasiteEntamoeba histolyticais car-ried out solely by the glycolytic pathway. Little kinetic and structural infor-mation is available for most of the pathway enzymes. We report here the gene cloning, overexpression and purification of hexokinase, hexose-6-phos-phate isomerase, inorganic pyrophosphate-dependent phosphofructokinase, fructose-1,6 bisphosphate aldolase (ALDO), triosephosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoglycerate kinase, phosphoglycerate mutase (PGAM), enolase, and pyruvate phos-phate dikinase (PPDK) enzymes fromE. histolytica