At present, the vast majority of China's freshwater aquaculture fish breeding is almost entirely based on artificial propagation, and less research has been done to the fertilized eggs hatching, especially in terms of circulating water hatching. In order to explore the effect of closed circulating water hatching system on the hatching performance of the fertilized eggs of acipenseriformes, in this paper, the hatching rate of the fertile eggs in acipenseriformes, the constitute and content of hatching enzyme, and the change situation of the concentration of ammonia nitrogen in the water body during the hatching process were studied. The results showed that the total concentration of ammonia nitrogen in the circulating water hatching system increased linearly with the increasing of hatch period; the ammonia excretion rate of the fertilized eggs in acipenseriformes was 60~90 mg/(g·h); the hatching rate was 86% in 6 hours after the rupture of membranes, and the accumulation of hatching enzymes in water body was serious. The hatching enzyme of acipenseriformes is mainly composed of the protein with molecular weight of 41.3 kDa. The hatching solution is composed of 23 kinds of proteins, including 7 kinds of hydrolases. The study showed that the removal of ammonia nitrogen and the hatching enzyme controlling technology are urgently to be resolved for the circulating water hatching system.