We used multienzyme coupled catalysis to produce L-Tyrosine(L-Tyr). In this study, Glycine was catalyzed to synthesize D-Serine by Aldolase(ALD), and D-Serine was transferred to pyruvic acid ammonium by D-Serine Dehydratase(SDH). Then, L-Tyr was synthesized by Tyrosine Phenol-Lyase(TPL) when the substrate phenol was added into the reaction system.The optimal reaction conditions were shown as follows: pH=8.5, 35 ℃, 6 g/L of ammonium acetate, 0.1 g/L of PLP, m(ALD)：m(SDH)：m(TPL)=6∶3∶5. We used the above conditions to catalyze glycine at the concentration of 50 g/L for 11 h, the concentration of L-Tyr was 117 g/L with a conversion of 97%, increasing by 4% compared with small-scale test and the yield reached 85%.The method of biosynthesis of L-Tyr can not only eliminate the purification process of D-Serine and pyruvic acid ammonium, but also has a reference significance for the reasonable utilization of resources and green synthesis process.