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(620x) Engineering Escherichia coli for Production of 3-Dehydroshikimate and Its Derivative Aromatic Compounds

Chen, W., Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences
Peng, Y., Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences
Wang, Q., Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences

3-dehydroshikimate (DHS) is a key metabolic intermediate for aromatic compound production. 3-dehydroshikimate (DHS) and its related aromatic compounds could be important feedstocks for producing a variety of important industrial chemicals, fibers and plastics with great commercial value. In the previous study, Escherichia coli was always engineered to produce DHS by deleting aroE and overexpressing several key genes via plasmid under the control of inducible promoters. This would result in the genetically instability of the engineered E coli and the additional supplement of minor shikimate or aromatic amino acids in the fermentation medium for cell growth, thus affect the future industrial production of DHS. Hence, it is greatly necessary to construct the genetically stable strain as well as adaptable to simple synthetic media with any supplementation for higher and efficient production of DHS. Here, in this study, the synthetic regulatory parts with different strength were used for combinatorially regulating the different modules of DHS synthetic pathway for facilitating glucose transport, enhancing and balancing the supply of precursors, and blocking the degradation of DHS. These modifications further increased the production of DHS in the simple mineral salt media without supplementing shikimate or aromatic acids by about 40 fold. The metabolically engineered strain produced about 50g/L DHS in 7-L fed-batch fermentation with more than 30% yield (mol/mol) from D-glucose. With this chassis cell for DHS production, many other aromatic compounds, such shikimate, galliate quinate, protocatechuate, catechol, vanillate and vanillin were efficiently  produced by redirect the flux based introducing heterogeneous enzymes. Our study laid important foundation for biomanufacturing of many useful aromatic compounds.