ENGINEERED REVERSAL OF THE ß-OXIDATION CYCLE FOR FUEL AND CHEMICAL PRODUCTION

We constructed a functional reversal of the b-oxidation cycle as a platform for the production of fuels and chemicals by engineering global regulators and eliminating native fermentative pathways (Nature 476, 355-359, 2011). However, this system-level approach makes it difficult to determine which of the many deregulated enzymes are responsible for product synthesis and limits efforts to fine-tune the synthesis of specific products and prevents the transfer of the engineered pathway to other organisms. To overcome these limitations, we have recently used a synthetic, bottom-up approach to construct and functionally characterize the three functional modules of the engineered b-oxidation reversal: i) priming, ii) elongation, and iii) termination (ACS Synth. Biol 1, 541-554, 2012). Each functional module was kinetically characterized in vitro followed by their in vivo assembly and functional characterization. In this talk, I will highlight our recent work in the use of this synthetic biology approach for the engineering of this pathway in Escherichia coli. I will also demonstrate how the modular nature of this metabolic platform allows its use for the synthesis of a wide array of functional molecules with applications in fuel and chemical production.