Abstract: Neutral phytase producing strains were screened by laser compound mutagenesis, mix wavelength 1.06+0.53 μm, irradiated time 1 min, irradiated power 15 w; The orientation test show that the produce phytase was extracellular-enzyme; Enzymology property mensurate show that E. Coli produce phytase optimal pH value was 7.4, optimal temperature was 37℃, B. subtilis produce phytase optimal pH value was 7.4, optimal temperature was 37℃; Enzymology property analysis show that E. Coli produce phytase has some extent pH flexibility and thermal endurance.