For the production of proteolytic enzyme wilth Aspergillus, the examination is made on the culture-time and koji extracting conditions, during producing koji. 1. The highest activity showed up when the culture-time took 50 hours for Aspergillus sojae and 60 hours for Aspergillus flavus. 2. When the cultured koji was extracted by a buffer solution and water, the former gave the product of higher activity until pH 7 through pH 12, and water until pH 3 through pH 7. 3. In the method of crushing and granule extractions, crushing extraction produced the one of higher activity than granule. 4. The highest activity showed up when Aspergillus sojae took 5 hours (Aspergillus flavus 4 hours) in the time of extracting enzyme solution. 5. The highest activity showed up when both Aspergillu sojae and Aspergillus flavus reacted and indicated $37.60^{circ}C$ in the reaction temperature and activity.