This study examines the effects of zinc amino acid (ZnAA) to the first sexual maturity stage in female broodstock of the Walking catfish, Clarias macrocephalus. The different ZnAA levels of Control (0 ppm ZnAA), ZnAA1 (100 ppm ZnAA) and ZnAA2 (200 ppm ZnAA) in the diet was applied to the first sexual maturation female catfish (Availa®Zn, Zinpro Corporation, Eden Prairie, MN USA). ZnAA accumulation, broodstock maturation analysis and breeding performance were evaluated. The ZnAA treatment has significant different in serum, meat and ovary ZnAA accumulation. The ZnAA treatment increased the fecundity, gonadosomatic index, egg diameter and development of oocytes at tertiary yolk stage. In comparison, the ZnAA treatment was insignificant in estradiol level. During artificial fertilization, the ZnAA treatment enhanced the fertilization rate and the larval survival rate. During recovery breeding, ZnAA treatment significantly increased the egg production and larval hatching rate. The optimum level to enhance the Clarias macrocephalus female broodstock first sexual maturation is ZnAA1.