Shikimate, a key metabolic intermediate in the biosynthesis of aromatic compounds in microorganisms and plants, is a valuable compound used as a starting material for industrial synthesis of an anti-influenza drug oseltamivir phosphate (Tamiflu). In this study, we constructed shikimate-overproducing Corynebacterium glutamicum strains by metabolic engineering and developed a highly efficient bioprocess for shikimate production by using growth-arrested high density cells of recombinant strains. Combinatorial effects of overexpression of shikimate pathway enzymes, inhibition of byproduct formation, improvement of precursor availability, and overexpression of a limiting glycolytic enzyme markedly enhanced shikimate productivity. The recombinant strain produced 141 g/l of shikimate from glucose at a yield of 51% (mol/mol) in minimal medium after 48h, representing the highest values for microbial shikimate production reported to date. Moreover, comparable high shikimate productivity could be achieved through simultaneous utilization of glucose, xylose, and arabinose, which would enable efficient utilization of lignocellulosic feedstocks.
