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Cephalosporium sp. KM 388 secreted protease into the culture broth after mycelial growth had ceased in complex medium, and concurrently with growth in semi-synthetic medium. Protease activity decreased rapidly when glucose was added to a culture in complex medium during the phase of enzyme accumulation. This decrease was proportional to the concentration of glucose added. After exhaustion of added glucose, the protease activity began to increase. Cycloheximide inhibited this increase, which suggests de novo synthesis of the protease. Addition of Polypepton also decreased protease activity in the complex medium. In contrast, glucose had no effect on the activity of protease produced in semi-synthetic medium. The effect of glucose in complex medium was reduced when mycelia were exposed to sodium nitrate before protease formation began. The decrease in protease activity stopped when mycelia were removed from the culture. In dialysis culture, glucose addition to the culture did not cause the decrease in protease activity even in the complex medium.