Furthermore, a new modular pathway from OSU6162 hydrochloride D-glucose to L-tyrosine was assembled and reoptimized with the identified optimal modules to enable de novo synthesis of -naringenin. The optimum strain was Phenamil methanesulfonate salt capable of producing 100.64 mg/L -naringenin from Dglucose, which is the highest production titer from D-glucose in Escherichia coli. The biosynthesis of flavonoids from safe, inexpensive, and renewable substrates is increasingly attracting attention due to concerns about food safety and environmetal issues. Previous studies have succeeded in producing -naringenin from p-coumaric acid. However, its high cost and poor water solubility restricted the direct application of phenylpropanoid acid precursors to industrial scale applications. In particular, these precursors are unfavorable commercially in terms of food safety issues because most of the chemicals are obtained by chemical synthesis routes through acetylsalicyloyl chloride from the petroleum industry. In this study, a bacterial platform for – naringenin production directly from D-glucose was constructed. The strategy described here would decrease substrate-related costs and facilitate the extensive application of -naringenin in both the pharmaceutical and nutraceutical industries. Previous studies have demonstrated the feasibility of de novo production of -naringenin. The engineered strain could produce 46 mg/L naringenin from D-glucose and up to 84 mg/L with the addition of the expensive fatty acid enzyme inhibitor, cerulenin. However, the expressions of genes were only examined individually as part of the overall pathway, which would constrain production of the desired compound due to imbalance in the overall pathway. In this study, the overall pathway, including the upstream pathway from D-glucose to L-tyrosine, the downstream pathway from L-tyrosine to – naringenin, and the malonate assimilation pathway, has been optimized. The optimal strain was capable of producing 100.64 mg/L -naringenin without the addition of cerulenin, which is the highest reported production titer from D-glucose in E. coli. This proves the necessity of varying the expressions of modules simultaneously.