The present invention relates to a mutant strain of Aspergillus sojae with an enhanced protease activity and a preparation method of a natural taste enhancer using the same. More specifically, the present invention relates to the mutant strain of Aspergillus sojae with an enhanced protease activity obtained by selecting a strain with high protease activity and treating it with N-methyl-N-Nitroso-N-nitrosoguanidine (NTG) and inducing a mutation through irradiation, and a preparation method of a natural taste enhancer using protein hydrolysate obtained by