This revised proposal is focused on three complementary specific aims: (1) development of an E. coli strain capable of supporting shikimic acid biosynthesis efficiently, while not resorting to the PTS system for sugar transport (which has poor atom economy), (2) Evaluate the pros and cons of engineering shikimic acid biosynthesis in S. cerevisiae and G. oxydans, neither of which rely on the PTS system for glucose transport, and (3) develop strategies for further buttressing shikimic acid biosynthesis by exogenous addition of simple feedstocks such as TCA cycle intermediates to cell cultures.
