The present study investigated the effect of the material type (defatted (DF) or non-defatted
(NDF)) combined with alcalase concentration (from 0.2 to 1.2%) and seven enzymatic hydrolysis
treatments on the enzymatic hydrolysis of snakehead fish head (SFH) using endoproteases and
exoproteases. After enzymatic hydrolysis, the fish protein hydrolysate was spray-dried, and the quality
of the hydrolysate (FPH) powder was investigated. The results revealed that the ideal material and
alcalase concentration were NDF and 0.8%, respectively. NDF was hydrolyzed using seven enzymatic
hydrolysis treatments: single (Alcalase (A), Protamex (P), and Flavourzyme (F)), simultaneous combined
(A + F, P + F), and sequential (A > F, P > F) treatments. FPH production using sequential (A > F)
treatment resulted in the highest total amino acid content, protein recovery, and degree of hydrolysis
among all enzymatic hydrolysis treatments. The FPH powder achieved good nutritional quality and had a
high protein content, full essential amino acids, and low lipid content. This, coupled with food safety with
no detected pathogenic bacteria and low heavy metal content in the FPH powder, were all below the
allowed limit. These results demonstrate that SFH can be used as a raw material to produce FPH powder
with high nutritional quality and food safety in the food industry