Abstract: Actinomucor elegans and Wutungkiao Mucor were used as experimental materials, through single factor and orthogonal experiments, the optimization of fermentation medium composition for high-yielding protease of two kinds of hair mould was researched. The results showed that the optimal fermentation medium composition for neutral protease of Actinomucor elegans was:sucrose content 10%, soybean flour 2.5%, KH2PO40.4%, CaCl20.11%, the neutral protease activity was 1310.56U/mL. The optimal fermentation medium composition for neutral protease of Wutungkiao Mucor was:sucrose content 10%, soybean flour 3.0%, KH2PO40.37%, CaCl20.07%, the last neutral protease activity was 1306.38U/mL. The improve percentage of neutral protease activities were 60.1% and 46.7% respectively. This research laid the foundation for follow-up study of shortenning the fermentation cycle of fermented bean curd and production process of direct-loading fermented bean curd.