In previous paper, it was shown that toxopyrimidine is phosphorylated by partially purified pyridoxal kinase in mouse brain to toxopyrimidine phosphate. In present report, the author tried to clarify whether the phosphorylations of pyridoxal and toxopyrimidine are catalysed by the same enzyme or not using highly purified preparation. It was concluded that the kinase is able to phosphorylate the both substrates since the heat stabilities, metal requirements, behaviors for inhibitors almost coincide in both activities.