Metabolic engineering of endogenous MEP pathway for enhanced lycopene production in Escherichia coli

Microbial cell factories represent the primary approach for heterologous lycopene synthesis, where gene source selection and pathway regulation have been demonstrated to have a significant impact on lycopene titer. In this study, key lycopene biosynthesis genes (<i>crtE</i>, <i>crtB</i> and <i>crtI</i>) derived from the extremophile <i>Deinococcus wulumuqiensis</i> R12 were introduced into <i>Escherichia coli</i>, generating the chassis strain H0. Fermentation optimization revealed sodium pyruvate significantly enhanced lycopene production and cell growth. Quantitative polymerase chain reaction (qPCR) analysis revealed that sodium pyruvate upregulated the expression of <i>dxr</i>, <i>ispA</i>, <i>crtE</i>, <i>crtB</i> and <i>crtI</i> genes, while downregulating the expression of <i>dxs</i> and <i>idi</i> genes. Consequently, different sources of <i>dxs</i>, <i>dxr</i>, <i>idi</i> and <i>ispA</i> genes were screened and co-expressed to reinforce the 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway in <i>E. coli</i>. The optimized combination of <i>dxs</i> from <i>E. coli</i> MG1655 with <i>idi</i> from <i>D. wulumuqiensis</i> R12 achieved maximal lycopene titer of 293.70 mg/L (112.49 mg/g DCW), which was 33.88-fold higher than that of the initial strain H0. This study offers genetic resources for heterologous carotenoid synthesis and establishes a reference framework for the synthesis of analogous complex isoprenoid metabolites.