Fig. 1.

Fig. 1. Pathway engineering strategy for shikimate production in C. glutamicum. (A) Shikimate metabolic pathway in C. glutamicum. The bold arrows and crosses indicate the steps for which corresponding genes were overexpressed and disrupted, individually. The dashed lines represent several catalytic steps. The genes involved in each step are shown in italics. G6P, glucose-6-phosphate; F6P, fructose-6-phosphate; GAP, glyceraldehyde-3-phosphate; PEP, phosphoenolpyruvate; PYR, pyruvate; AcCoA, acetyl-CoA; OAA, oxaloacetate; Ru5P, ribulose-5-phosphate; X5P, xylulose-5-phosphate; R5P, ribose- 5-phosphate; S7P, sedoheptulose-7-phosphate; E4P, erythrose-4-phosphate; DHAP, 3-deoxy-D-arabinoheptulosanate-7- phosphate; DHQ, 3-dehydroquinate; DHS, 3-dehydroshikimate; PCA, protocatechuate. Genes and corresponding enzymes are as follows: pyk1, pyruvate kinase 1; aroF and aroG, DAHP synthase; aroB, 3-dehydroquinate synthase; qsuC, dehydroquinate dehydratase; aroE, shikimate dehydrogenase; aroK, shikimate kinase; qsuD, quinate/shikimate dehydrogenase; qsuB, dehydroshikimate dehydratase. (B) Gene disruption in C. glutamicum ATCC13032 and plasmid construction. The constructed plasmids were introduced and replicated in Inha304, yielding Inha305, Inha306, Inha307, Inha308, Inha309, and Inha310, respectively. CaroE and EaroE, shikimate dehydrogenase from C. glutamicum ATCC13032 and E. coli K-12, respectively; aroF^S188C, DAHP synthase carrying S188C mutation; EaroG^S180F, DAHP synthase carrying S180F mutation.

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