L-tyrosine is a value-added chemical in the food, pharmaceutical, chemical. Several researches to enhance Ltyrosine production have been implemented, but regulation of translation-level expression and carbon flux rebalancing around phosphoenolpyruvate (PEP) nod still has proven to be achieved for balancing the pathway. Here, we regulated metabolic pathway to enhance L-tyrosine production in Escherichia coli by fine-tuning of gene expression levels. To enhance the L-tyrosine production, a synthetic constitutive promoter and a synthetic 5’-UTR were designed for each gene of interest on the chromosome to control expression levels at both transcription and translational sides. Optimization of carbon flux was achieved by modulating the expression level of PEP synthase using UTR Designer. The L-tyrosine productivity of the modified E. coli strain, SCK5, was enhanced through pathway optimization resulting in 3.0 g/L of L-tyrosine titer increase.