Abstract: The optimum enzymatic hydrolysis conditions of Hippoglossus hippoglossus bone and amino acid composition characteristics of its enzymatic hydrolysate were studied in this paper. Based on the result of single factor experiments,response surface methodology(RSM)was carried out by hydrolysis degree. The results showed that the best enzymatic hydrolysis conditions were liquid ratio 2:1,enzyme addition 0.16%,enzymatic hydrolysis time 2 h. Under the condition,the degree of enzyme hydrolysis of Hippoglossus hippoglossus bone was 32.79%±0.33%. There were 8 essential amino acids required by the human body in the Hippoglossus hippoglossus bone enzymatic solution,which contained 33.3%. The content of some of the essential amino acids in Hippoglossus hippoglossus bone enzymatic solution and the essential amino acid content of milk protein and the ideal protein proposed by FAO/WHO were basically similar. It was indicated that the nutritional value of the Hippoglossus hippoglossus bone was higher. The enzymatic solution of Hippoglossus hippoglossus bone was rich in taste amino acids,with an effective amino acid content of 34.2%,a sweet amino acid content of 41.4%,a bitter amino acid content of 38.2%,and an umami amino acid content of 52.6%. The results showed that the enzymatic solution of Hippoglossus hippoglossus bone was of high nutritional value and good flavor. It could be used as a source of high quality protein or as a flavor modifier for specialty foods and other functional foods,as well as a research and development base for fresh seasonings. It was a marine protein resource with high application value and potential for development.