Biosynthetic pathways for the microbial production of hydroxycinnamoyl anthranilates from glucose. (A) Three compatible plasmids were used for the biological production of Avn D and Avn F from glucose. A first plasmid is a shikimate module which contains all the six genes for the production of shikimate (SHIK) from pyruvate (PYR), fructose 6-phosphate (F6P), and glyceraldehydes 3-phosphate (G3P) (orange box). A second plasmid is a tyrosine module which contains all the five genes for the production of tyrosine (TYR) from SHIK (green box). The third plasmid is the cinnamoyl anthranilate module which contains up to five genes for the conversion of TYR into two hydroxycinnamoyl-CoAs and their coupling to anthranilate (ANT) for the production of Avn D and Avn F (blue box).CAFA, caffeate; CAF-CoA, caffeoyl-CoA; CHA, chorismate;COUA, p-coumarate; COU-CoA, p-coumaroyl-CoA; DAHP, 3-deoxy-D-arabino-heptulosonate; DHQ, dehydroquinate; DHS, dehydroshikimate; E4P, erythrose 4-phosphate; EPSP, 5-enolpyruvoylshikimate 3-phosphate; HPP, 4 hydroxyphenlypyruvate; PEP, phosphoenolpyruvate; PPA, prephenate; and S3P, shikimate 3-phosphate. The enzymes (in bold face) are as follows: PpsA, phosphoenolpyruvate synthase; TktA, transketolase A; AroG, DAHP synthase; AroB, DHQ synthase; AroD, DHQ dehydratase; YdiB, shikimate dehydrogenase; AroL, shikimate kinase II; AroA, EPSP synthase; AroC, chorismate synthase; TrpE/D, anthranilate synthase I/II; TyrA, chorismatemutase/prephenate dehydrogenase; and TyrB, tyrosine aminotransferase; TAL, tyrosine ammonia-lyase; 4CL, p-coumarate:CoA ligase; COUA3H, p-coumarate 3-hydroxylase; HCBT, hydroxycinnamoyl/benzoyl-CoA/anthranilate N-hydroxycinnamoyl/benzoyltransferase. The dashed lines indicate where feedback inhibitions occur. Allosteric regulation of AroG and TyrA were removed by employing their respective feedback-resistant mutants, AroG* (D146N) and TyrA* (M53I; A354V), respectively. (B) Structures of the three plasmids used for cinnamoyl anthranilates production. The open blocks indicate the origins of replication, the open arrows represent the genes, and the angled arrows indicate the promoters.