STARTUP - Sustainable Aromatics Using Pseudomonas
Final Report Abstract
Aromatic chemicals are a cornerstone of modern society, with more than 40 million tons per year produced as building blocks for the production of plastics, pharmaceuticals, food ingredients and many other applications. Currently, the vast majority of these chemicals are produced from petroleum. However, the fear of depletion of this important resource is ever increasing, and the number of oil producing countries is small and these countries are often politically unstable. These factors contribute to large fluctuations in the price of oil, which are reflected by the prices of aromatics. Also, the chemical production of aromatics involves toxic solvents and catalysts, high temperature and pressure, hazardous and polluting waste and it contributes greatly to global warming through greenhouse gas emissions. To address these problems, the STARTUP projects aims to produce aromatics chemicals such as phenol and p-hydroxybenzoate from renewable resources. Biological production of aromatics is potentially much cleaner, since it is performed at ambient temperature and pressure, produces mostly biodegradable waste, and lacks many of the emissions commonly associated with the petrochemical industry (NOx, SO2, VOC). Most importantly, since bio-chemicals are produced from biomass-derived substrates (e.g. sugar, glycerol), it greatly reduces net CO2 emissions (closed carbon cycle), leading to a much more sustainable process. The STARTUP project has made significant advances in the production of aromatic chemicals from sugars. We use a solvent-tolerant Pseudomonas bacterium, which is uniquely capable of growing in the presence of aromatics that are deadly to other bacteria. Through metabolic engineering, we have enabled Pseudomonas to produce a wide range of aromatics with applications ranging from bulk polymers to specialty pharmaceuticals. We have also optimized the production of these aromatics by fundamentally altering the metabolism of the organism. In doing so, we have learned much about how and why Pseudomonas produces, and degrades, aromatic chemicals. We can now apply this knowledge to make new biocatalysts for other aromatic products. Surprisingly, with the right knowledge, only very small genetic modifications needed to be made in the organism in order to get it to produce aromatic chemicals. In addition, we have streamlined the organism itself to grow more efficiently in the highly artificial environment of a bioreactor, thereby facilitating industrial applications. We have also contributed new synthetic biology tools to the Pseudomonas community, contributing to the overall scientific research of this organism. In the future, we expect to further develop the biocatalysts for other substrates and products, and to gain a deeper understanding of their inner workings. Doing so, we hope to contribute to the establishment of a circular, bio-based economy, which would be of great benefit to society, economy and environment.
Publications
- 2015. Complete genome sequence of solvent-tolerant Pseudomonas putida S12 including megaplasmid pTTS12. J. Biotechnol. 200:17-18
Küpper, J., H.J. Ruijssenaars, L.M. Blank, J.H. de Winde, N. Wierckx
(See online at https://doi.org/10.1016/j.jbiotec.2015.02.027) - 2015. Engineering mediatorbased electroactivity in the obligate aerobic bacterium Pseudomonas putida KT2440. Front. Microbiol. 6:284
Schmitz, S., S. Nies, N. Wierckx, L.M. Blank, M.A. Rosenbaum
(See online at https://doi.org/10.3389/fmicb.2015.00284) - 2015. Metabolic engineering of Pseudomonas putida KT2440 to produce anthranilate from glucose. Front. Microbiol. 6:1310
Küpper, J., J. Dickler, M. Biggel, S. Behnken, G Jaeger, N. Wierckx, L.M. Blank
(See online at https://doi.org/10.3389/fmicb.2015.01310) - 2015. Tn7-based device for calibrated heterologous gene expression in Pseudomonas putida. ACS Synth. Biol. 4:1341-1351
Zobel, S., I. Benedetti, L. Eisenbach, V. de Lorenzo§, N. Wierckx, L.M. Blank
(See online at https://doi.org/10.1021/acssynbio.5b00058) - 2016. Regulation of solvent-tolerance in Pseudomonas putida S12 mediated by mobile elements. Microb. Biotechnol. 10:1558-1568
Hosseini, R., J. Kuepper, S. Koebbing, L.M. Blank, N. Wierckx, J.H. de Winde
(See online at https://doi.org/10.1111/1751-7915.12495) - 2017. Metabolic response of Pseudomonas putida to increased NADH regeneration rates. Eng. Life Sci. 17(1):47-57
Zobel, S., J. Kuepper, B. Ebert, N. Wierckx, L.M. Blank
(See online at https://doi.org/10.1002/elsc.201600072) - 2018. Engineering Pseudomonas putida KT2440 for efficient ethylene glycol utilization. Metab. Eng. 48:197-207
Franden, M.A., L. Jayakody, W.-J. Li, N.J. Wagner, N.S. Cleveland, W.E. Michener, B. Hauer, L.M. Blank, N. Wierckx, J. Klebensberger, G.T. Beckham
(See online at https://doi.org/10.1016/j.ymben.2018.06.003) - 2018. Metabolic engineering of Pseudomonas taiwanensis VLB120 with minimal genomic modifications for highyield phenol production. Metab. Eng. 47:121-133
Wynands, B., C. Lenzen, M. Otto, F. Koch, L.M. Blank, N. Wierckx
(See online at https://doi.org/10.1016/j.ymben.2018.03.011) - 2019. High-yield production of 4-hydroxybenzoate from glucose or glycerol by an engineered Pseudomonas taiwanensis VLB120. Front. Bioeng. Biotechnol.
Lenzen, C., B. Wynands, M. Otto, J. Bolzenius, P. Mennicken, L.M. Blank, N. Wierckx
(See online at https://doi.org/10.3389/fbioe.2019.00130) - 2019. Laboratory evolution reveals the metabolic and regulatory basis of ethylene glycol metabolism by Pseudomonas putida KT2440. Environ. Microbiol.
Li, W.-J., L.N. Jayakody, M.A. Franden, M. Wehrmann, T. Daun, B. Hauer, L.M. Blank, , G.T. Beckham, J. Klebensberger, N. Wierckx
(See online at https://doi.org/10.1111/1462-2920.14703) - 2019. Pseudomonas putida rDNA is a favored site for the expression of biosynthetic genes. Sci Rep. 9:7028
Domröse, A., J. Hage-Hülsemann, S. Thies, R. Weihmann, L. Kruse, M. Otto, N. Wierckx, K.- E. Jaeger, T. Drepper, A. Loeschcke
(See online at https://doi.org/10.1038/s41598-019-43405-1) - 2019. Rational engineering of L-phenylalanine accumulation in Pseudomonas taiwanensis to enable high-yield production of trans-cinnamate. Front. Bioeng. Biotechnol.
Otto, M., B. Wynands, C. Lenzen, M. Filbig, L.M. Blank, N. Wierckx
(See online at https://doi.org/10.3389/fbioe.2019.00312) - 2019. Streamlined Pseudomonas taiwanensis VLB120 chassis strains with improved bioprocess features. ACS Synth. Biol.
Wynands, B., M. Otto, N. Runge, S. Preckel, T. Polen, L.M. Blank, N. Wierckx
(See online at https://doi.org/10.1021/acssynbio.9b00108) - 2019. Targeting 16S ribosomal DNA for stable recombinant gene expression in Pseudomonas. ACS Synth. Biol.
Otto, M., B. Wynands, T. Drepper, K.-E. Jaeger, S. Thies, A. Loeschcke, L.M. Blank, N. Wierckx
(See online at https://doi.org/10.1021/acssynbio.9b00195)
