To provide methods for effectively producing 1,4-butanediol (BOD) and chemical precursors thereof in commercial quantities.SOLUTION: Disclosed herein is a non-naturally occurring microorganism comprising at least one exogenous nucleic acid encoding an enzyme in BOD production pathway to express the exogenous enzyme with a disrupted gene encoding an aerobic respiratory control regulatory system. The exogenous enzymes are succinyl-CoA synthetase, alpha-ketoglutarate decarboxylase, succinate semialdehyde dehydrogenase and 4-hydroxybutyrate dehydrogenase and optionally 4-hydroxybutyryl-CoA/acetyl-CoA transferase, butyrate kinase and phosphotransbutyrase, 4-hydroxybutyryl-CoA reductase, 4-hydroxybutanal reductase, pyruvate dehydrogenase, NADH-insensitive citrate synthetase and phosphoenolpyruvate carboxykinase.SELECTED DRAWING: Figure 46

Genomatica has established an integrated computational/experimental metabolic engineering platform to design, create, and optimize novel high performance organisms and bioprocesses. Here we present our platform and its use to develop E. coli strains for production of the industrial chemical 1,4-butanediol (BDO) from sugars. A series of examples are given to demonstrate how a rational approach to strain engineering, including carefully designed diagnostic experiments, provided critical insights about pathway bottlenecks, byproducts, expression balancing, and commercial robustness, leading to a superior BDO production strain and process.