We have developed a functional reversal of the �-oxidation cycle as an efficient pathway for the production of fuels and chemicals by engineering global regulators and eliminating native fermentative pathways in Escherichia coli (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 recently used a synthetic, bottom-up approach to construct and functionally characterize the three functional modules of the engineered �-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 E. coli. I will also demonstrate how the modular nature of this metabolic platform enables the synthesis of a wide array of functional molecules with applications in fuel and chemical production.

Modular biosynthesis for the production of advanced fuels and chemicals

247th National Meeting (2014)

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