ABSTRACT

It is a national priority to accelerate biomanufacturing and to reduce dependence on refining petroleum to obtain commodity chemicals. This project focuses on developing techniques and bacterial strains that will allow industrially important compounds to be made from renewable resources rather than from fossil fuels. Towards this goal, the ability of bacteria to synthesize complex chemicals can be exploited. Among the petrochemicals in high demand are aromatic compounds that can be made by bacteria. However, natural biosynthetic pathways need to be modified to allow the accumulation of specific compounds at high levels to be economically feasible. This project involves the modification of a biosynthetic pathway in two kinds of soil bacteria, Acinetobacter baylyi and Pseudomonas putida to produce aromatic compounds. The research is accomplished through a collaboration with a team of scientists at Argonne National Laboratory. This collaboration has both scientific and educational components. Students learn about biomanufacturing and metabolic engineering, as well as careers in these fields, through a seminar series featuring scientists of the Agile BioFoundry.

The long-term goal of this research is to create a cost-effective biosynthetic route for the bacterial conversion of plant-derived sugars to industrially needed aromatic compounds. Such compounds are in high demand and are commercially available now as petrochemicals. The bacteria employed in the research, Acinetobacter baylyi ADP1 and Pseudomonas putida KT2440, use the shikimate pathway to make aromatic compounds. To increase carbon flux into biosynthesis, an enzyme that is not typically part of the shikimate pathway is altered by novel mutagenesis and adaptive laboratory evolution methods and investigated in A. baylyi. This bacterium has an exceptionally high efficiency of natural transformation and allelic replacement and serves as host to generate novel enzymes. A modified shikimate pathway is engineered in P. putida using one of the novel enzymes generated in A. baylyi. This strategy uses the two different kinds of bacteria in a complementary approach that exploits unique features of each. The engineered pathway in P. putida is predicted to redirect carbon from a renewable biomass feedstock into high levels of desired end products. The collaboration with researchers from the Agile BioFoundry employs high throughput techniques to accomplish the project goals using advanced analytics and microfluidics-based techniques.

This award is co-funded by the Systems and Synthetic Biology program in the Division of Molecular and Cellular Biosciences and the Cellular and Biochemical Engineering program in the Division of Chemical Bioengineering Environmental and Transport Systems.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

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