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000874551 1001_ $$0P:(DE-Juel1)176879$$aLi, Wing-Jin$$b0
000874551 245__ $$aUnraveling 1,4-Butanediol Metabolism in Pseudomonas putida KT2440
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000874551 520__ $$aPlastics, 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.
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000874551 7001_ $$0P:(DE-HGF)0$$aNarancic, Tanja$$b1
000874551 7001_ $$0P:(DE-HGF)0$$aKenny, Shane T.$$b2
000874551 7001_ $$0P:(DE-HGF)0$$aNiehoff, Paul-Joachim$$b3
000874551 7001_ $$0P:(DE-HGF)0$$aO’Connor, Kevin$$b4
000874551 7001_ $$0P:(DE-HGF)0$$aBlank, Lars M.$$b5
000874551 7001_ $$0P:(DE-Juel1)176653$$aWierckx, Nick$$b6$$eCorresponding author$$ufzj
000874551 773__ $$0PERI:(DE-600)2587354-4$$a10.3389/fmicb.2020.00382$$gVol. 11, p. 382$$p382$$tFrontiers in microbiology$$v11$$x1664-302X$$y2020
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