(15g) Metabolic Engineering of Bacteria for Production of Oleochemicals

Pfleger, B., Chemical & Biological Engineering, University of Wisconsin, Madison, WI
Finding a sustainable alternative for today’s petrochemical industry is a major challenge facing chemical engineers and society at large. To be sustainable, routes for converting carbon dioxide and light into organic compounds for use as both fuels and chemical building blocks must be identified, understood, and engineered. Advances in metabolic engineering, synthetic biology, and other bioengineering disciplines have expanded the scope of what can be produced in a living organism. As in other engineering disciplines, synthetic biologists want to apply a general understanding of science (e.g. microbiology and biochemistry) to construct complex systems from well-characterized parts (e.g. DNA and protein). Once novel synthetic biological systems (e.g. enzymes for biofuel synthesis) are constructed, they must be engineered to function inside evolving cells without negatively impacting the host’s physiology.

In this talk, I will describe pathways for producing high-value commodity chemicals derived from fatty-acids and how my group and others have combined synthetic biology and systems biology to improve oleochemical production in bacteria using sustainable feedstocks. The talk will describe the critical regulatory points in native fatty acid metabolism, strategies for deregulating the pathway, and alternatives that by-pass it altogether. I will highlight the use of heterologous plant and bacterial enzymes to alter the chain length distribution of products from common long-chain molecules to higher-value medium-chain analogs. I will also describe our collaborative efforts with the Maranas lab at Penn State to use computational tools to redesign the substrate specificity of enzymes used to produce medium chain oleochemicals. Here, we have used the IPRO software to identify a small (~60) library of thioesterase mutants that resulted in a new variant that produces 2-fold more fatty acid with over 50% C8 chain lengths, molecules not produced by the native enzyme.