Alkaline phosphatase activity appeared in Streptomyces noursei strain IMET 43,716 when cultures were shifted to phosphate limitation. By using p-nitrophenylphosphate as substrate, the activity detected at pH 9.4 was cell-bound, as long as young mycelia were studied. Supernatant fluids of the cultures were only active, if partial mycelial lysis had taken place after incubation for several days under the influence of shear forces. After cytochemical staining the phosphatase reaction products were microscopically visible as grains distributed rather homogeneously within the hyphal lumen. The frequency of grains was correlated to the rate of nourseothricin production. Electron microscopy of thin sections showed the phosphatase reaction products to be only present in the cytoplasm.