Carbamyl phosphate synthesis in Bacillus subtilis.
In vitro and "in situ" assays have been developed to test the carbamyl phosphate synthetase (CPSase) activity of a series of pyrimidine-requiring mutants of Bacillus subtilis. The enzyme has been shown to be highly unstable, and was successfully extracted only in the presence of 10% glycerol and 1 mM dithiothreitol (Cleland's reagent). It loses activity rapidly when sonicated or when treated with lysozyme. Genetic studies, using mutants, indicate that B. subtilis may possess two CPSases. This possibility and its physiological consequences were probed enzymatically. CPSase activity has been shown to undergo inhibition by both uridine triphosphate and dihydroorotate; activation has been demonstrated in response to phosphoribosyl pyrophosphate (PRPP) and (to a lesser extent) ornithine.
D001412:Bacillus subtilis / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D002219:Carbamates / Q000096:biosynthesis*; D002221:Carbamyl Phosphate / Q000096:biosynthesis*; D002501:Centrifugation, Zonal; D002850:Chromatography, Gel; D004269:DNA, Bacterial / Q000032:analysis; D004789:Enzyme Activation; D005973:Glutamine / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D009113:Muramidase / Q000494:pharmacology; D009154:Mutation; D010754:Phosphoribosyl Pyrophosphate / Q000494:pharmacology; D010770:Phosphotransferases / Q000378:metabolism*; D011312:Pressure; D011743:Pyrimidines / Q000378:metabolism; D013010:Sonication; D014170:Transformation, Genetic
D002219:Carbamates; D004269:DNA, Bacterial; D011743:Pyrimidines; D005973:Glutamine; D002221:Carbamyl Phosphate; D010754:Phosphoribosyl Pyrophosphate; D010770:Phosphotransferases; D009113:Muramidase