Metabolic flux analysis for Escherichia coli by flux balance analysis.

Conventional metabolic flux analysis (MFA) of Escherichia coli wild type and of pathway gene knockout mutants cultivated under anaerobic condition is explained in detail in this chapter. To place the MFA results into the context of the literature, the regulation of central carbon metabolism in terms of catabolite regulation by the phosphotransferase system (PTS) and the response to oxygen limitations via global regulators is reviewed. The effects of gene deletions such as pflA, pta, ppc, pykF, adhE, and ldhA on the metabolic network are presented. Moreover, for the pflA mutant the effects of various carbon sources were quantified. The chapter thereby contributes to the discussion of metabolic network function and the design of microbial cell factories.