Abstract
To drive innovation in chemical and material applications beyond what has been afforded by the mature petrochemical industry, new molecules that possess diverse chemical functionality are needed. One source of such molecules lies in the varied metabolic pathways that soil microbes utilize to catabolize aromatic compounds generated during plant decomposition. Here, we have engineered Pseudomonas putida KT2440 to convert these aromatic compounds to 15 catabolic intermediates that exhibit substantial chemical diversity. Bioreactor cultivations, analytical methods, and bench-scale separations were developed to enable production (up to 58 g/L), detection, and purification of each target molecule. We further engineered strains for production of a subset of these molecules from glucose, achieving a 41% molar yield of muconic acid. Finally, we produce materials from three compounds to illustrate the potential for realizing performance-advantaged properties relative to petroleum-derived analogs. In the last century, chemicals and materials derived from the byproducts of petroleum production have largely displaced natural products and enabled myriad new applications. Today, fuels, chemicals, and materials derived from plant biomass have the potential to enable innovation and mitigate negative environmental impacts of the petrochemical industry. To achieve this, nature's ability to generate unique molecules with great selectivity could be leveraged to develop new chemicals and materials that would be difficult to access from petroleum and could represent new building blocks for a bio-based materials economy. Here we describe the production of molecules derived from bacterial aromatic catabolic pathways and demonstrate their use in the production of materials with superior properties relative to their petroleum-derived analogs. Intermediates of bacterial aromatic catabolism contain chemical functionality that could enable them to serve as precursors to environmentally compatible materials with similar or superior properties relative to petroleum-derived incumbents. Here, Pseudomonas putida was engineered to convert aromatic molecules and glucose into 16 of these metabolic intermediates including muconic acid, which was produced at a 41% yield from glucose. Several of these molecules were then polymerized to generate performance-advantaged materials.
Original language | English |
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Pages (from-to) | 1523-1537 |
Number of pages | 15 |
Journal | Joule |
Volume | 3 |
Issue number | 6 |
DOIs | |
State | Published - Jun 19 2019 |
Funding
This work was partially authored by Alliance for Sustainable Energy, LLC, the manager and operator of the National Renewable Energy Laboratory for the U.S. Department of Energy under Contract No. DE-AC36-08GO28308. Oak Ridge National Laboratory is managed by UT-Battelle, LLC, for the U.S. DOE under contract DE-AC05-00OR22725. We acknowledge funding from NREL Laboratory Directed Research and Development (LDRD) program. Support is also acknowledged from the U.S. Department of Energy, Office of Energy Efficiency and Renewable Energy, Bioenergy Technologies Office via the Agile BioFoundry project for the development of strains CJ442 and CJ522 and techno-economic analysis. G.T.B. acknowledges partial support from the Center for Bioenergy Innovation, a U.S. Department of Energy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science. Support for N.A.R. was partially provided by the U.S. Department of Energy, Office of Science, Office of Workforce Development for Teachers and Scientists, Office of Science Graduate Student Research (SCGSR) program. The SCGSR program is administered by the Oak Ridge Institute for Science and Education for the DOE under contract number DE-AC05-06OR23100. Funding for C.R.M. was provided by the Colorado College Riley Scholar-in-Residence program for research support. We thank Sandra Notonier for use of her plasmid, pSN2, in strain construction and Eric Karp for helpful discussions on separations. C.W.J. D.R.V. and G.T.B. conceived of the project. C.W.J. G.D. J.R.E. P.K. and A.M.G. conducted the metabolic engineering. D.S. P.S. T.A.V. and X.Y. conducted the bioreactor cultivations. N.A.R. and C.R.M. conducted the separations. N.A.R. C.R.M. and A.N.W. performed the polymer synthesis. B.A.B. N.S.C. W.E.M. D.J.P. and K.J.R. conducted the analytics. D.R.V. N.G. and M.J.B. conducted the techno-economic analysis. P.C.S.J. and Y.J.B. performed the metabolic modeling. C.W.J. D.S. N.A.R. B.A.B. D.R.V. P.C.S.J. and G.T.B. wrote the manuscript with critical input from all authors. C.W.J. D.S. P.C.S.J. D.R.V. J.R.E. A.M.G. and G.T.B. hold patents and have submitted patent applications on engineered strains related to this work. N.A.R. D.R.V. and G.T.B. have submitted patent applications on the production of polymers from these molecules. This work was partially authored by Alliance for Sustainable Energy, LLC, the manager and operator of the National Renewable Energy Laboratory for the U.S. Department of Energy under Contract No. DE-AC36-08GO28308. Oak Ridge National Laboratory is managed by UT-Battelle, LLC, for the U.S. DOE under contract DE-AC05-00OR22725. We acknowledge funding from NREL Laboratory Directed Research and Development (LDRD) program. Support is also acknowledged from the U.S. Department of Energy, Office of Energy Efficiency and Renewable Energy , Bioenergy Technologies Office via the Agile BioFoundry project for the development of strains CJ442 and CJ522 and techno-economic analysis. G.T.B. acknowledges partial support from the Center for Bioenergy Innovation , a U.S. Department of Energy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science. Support for N.A.R. was partially provided by the U.S. Department of Energy, Office of Science, Office of Workforce Development for Teachers and Scientists, Office of Science Graduate Student Research (SCGSR) program. The SCGSR program is administered by the Oak Ridge Institute for Science and Education for the DOE under contract number DE-AC05-06OR23100. Funding for C.R.M. was provided by the Colorado College Riley Scholar-in-Residence program for research support. We thank Sandra Notonier for use of her plasmid, pSN2, in strain construction and Eric Karp for helpful discussions on separations.
Funders | Funder number |
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Colorado College Riley Scholar-in-Residence | |
DOE Office of Science | |
Office of Biological and Environmental Research | |
Office of Science Graduate Student Research | |
SCGSR | |
U.S. Department of Energy Research Center | |
U.S. Department of Energy | DE-AC05-00OR22725 |
Office of Science | |
Office of Energy Efficiency and Renewable Energy | CJ522 |
Workforce Development for Teachers and Scientists | |
Oak Ridge National Laboratory | |
Oak Ridge Institute for Science and Education | DE-AC05-06OR23100 |
National Renewable Energy Laboratory | |
Laboratory Directed Research and Development | |
Bioenergy Technologies Office | |
Center for Bioenergy Innovation |
Keywords
- aromatic catabolism
- biopolymer
- bioprocess development
- functional replacement
- muconic acid