We propose that amino acids in natal stream water have important roles in Pacific salmon homing. This study hypothesized that amino acids found in natal stream water have a role in the ability of mature male chum salmon (Oncorhynchus keta) to home to the Osaru River (OR), Hokkaido, Japan. Behavioral experiments were conducted in a two-choice test tank using various combinations of control water (natural Toya Lake water; NLW and three artificial stream waters using amino acids: 1) artificial OR water (AOR); 2) AOR without L-glutamic acid, the major amino acid in OR water (AOR-E); and 3) artificial water matching another stream (ALS) that had much higher amino acid concentrations than OR. In behavioral tests, the fish did not select between AOR and AOR-E, but still chose AOR over NLW, AOR-E over NLW, and AOR over ALS. These results suggest that migratory male chum salmon respond to amino acid mixtures in their natal stream water and appear to be affected by multiple amino acids.