This title appears in the Scientific Report :
2020
Please use the identifier:
http://hdl.handle.net/2128/25058 in citations.
Please use the identifier: http://dx.doi.org/10.3389/fmicb.2020.00382 in citations.
Unraveling 1,4-Butanediol Metabolism in Pseudomonas putida KT2440
Unraveling 1,4-Butanediol Metabolism in Pseudomonas putida KT2440
Plastics, in all forms, are a ubiquitous cornerstone of modern civilization. Although humanity undoubtedly benefits from the versatility and durability of plastics, they also cause a tremendous burden for the environment. Bio-upcycling is a promising approach to reduce this burden, especially for po...
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Personal Name(s): | Li, Wing-Jin |
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Narancic, Tanja / Kenny, Shane T. / Niehoff, Paul-Joachim / O’Connor, Kevin / Blank, Lars M. / Wierckx, Nick (Corresponding author) | |
Contributing Institute: |
Biotechnologie; IBG-1 |
Published in: | Frontiers in microbiology, 11 (2020) S. 382 |
Imprint: |
Lausanne
Frontiers Media
2020
|
PubMed ID: |
32256468 |
DOI: |
10.3389/fmicb.2020.00382 |
Document Type: |
Journal Article |
Research Program: |
Biotechnology |
Link: |
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Publikationsportal JuSER |
Please use the identifier: http://dx.doi.org/10.3389/fmicb.2020.00382 in citations.
Plastics, in all forms, are a ubiquitous cornerstone of modern civilization. Although humanity undoubtedly benefits from the versatility and durability of plastics, they also cause a tremendous burden for the environment. Bio-upcycling is a promising approach to reduce this burden, especially for polymers that are currently not amenable to mechanical recycling. Wildtype P. putida KT2440 is able to grow on 1,4-butanediol as sole carbon source, but only very slowly. Adaptive laboratory evolution (ALE) led to the isolation of several strains with significantly enhanced growth rate and yield. Genome re-sequencing and proteomic analysis were applied to characterize the genomic and metabolic basis of efficient 1,4-butanediol metabolism. Initially, 1,4-butanediol is oxidized to 4-hydroxybutyrate, in which the highly expressed dehydrogenase enzymes encoded within the PP_2674-2680 ped gene cluster play an essential role. The resulting 4-hydroxybutyrate can be metabolized through three possible pathways: (i) oxidation to succinate, (ii) CoA activation and subsequent oxidation to succinyl-CoA, and (iii) beta oxidation to glycolyl-CoA and acetyl-CoA. The evolved strains were both mutated in a transcriptional regulator (PP_2046) of an operon encoding both beta-oxidation related genes and an alcohol dehydrogenase. When either the regulator or the alcohol dehydrogenase is deleted, no 1,4-butanediol uptake or growth could be detected. Using a reverse engineering approach, PP_2046 was replaced by a synthetic promotor (14g) to overexpress the downstream operon (PP_2047-2051), thereby enhancing growth on 1,4-butanediol. This work provides a deeper understanding of microbial 1,4-butanediol metabolism in P. putida, which is also expandable to other aliphatic alpha-omega diols. It enables the more efficient metabolism of these diols, thereby enabling biotechnological valorization of plastic monomers in a bio-upcycling approach. |