Fig 2.
(A) Stepwise improvements of the shikimate module by changing the origin of replication from pSC101 to pBBR1 and the promoter from PLtetO-1 to Plac-UV5 (S1), followed by codon optimization of aroB (S2), replacement of ydiB with aroE (S3), and inserting a second promoter PLtetO-1 5′ of aroG* (S4). In S5, a combination of the rrnB terminator T1 (large T) and Ptrc was used to substitute PLtetO-1 5′ of aroG*, which resulted in significant reductions in protein and shikimate production. (B) The SRM results indicate the relative levels of TktA through YdiB/AroE as a consequence of the various modifications to the shikimate module. All cultures were performed under the conditions described in Table 3.