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@ARTICLE{Ackermann:892434,
author = {Ackermann, Yannic S. and Li, Wing-Jin and Op de Hipt,
Leonie and Niehoff, Paul-Joachim and Casey, William and
Polen, Tino and Köbbing, Sebastian and Ballerstedt, Hendrik
and Wynands, Benedikt and O'Connor, Kevin and Blank, Lars M.
and Wierckx, Nick},
title = {{E}ngineering adipic acid metabolism in {P}seudomonas
putida},
journal = {Metabolic engineering},
volume = {67},
issn = {1096-7176},
address = {Orlando, Fla.},
publisher = {Academic Press},
reportid = {FZJ-2021-02080},
pages = {29-40},
year = {2021},
note = {Biotechnologie 1},
abstract = {Bio-upcycling of plastics is an upcoming alternative
approach for the valorization of diverse polymer waste
streams that are too contaminated for traditional recycling
technologies. Adipic acid and other medium-chain-length
dicarboxylates are key components of many plastics including
polyamides, polyesters, and polyurethanes. This study endows
Pseudomonas putida KT2440 with efficient metabolism of these
dicarboxylates. The dcaAKIJP genes from Acinetobacter
baylyi, encoding initial uptake and activation steps for
dicarboxylates, were heterologously expressed. Genomic
integration of these dca genes proved to be a key factor in
efficient and reliable expression. In spite of this,
adaptive laboratory evolution was needed to connect these
initial steps to the native metabolism of P. putida, thereby
enabling growth on adipate as sole carbon source. Genome
sequencing of evolved strains revealed a central role of a
paa gene cluster, which encodes parts of the phenylacetate
metabolic degradation pathway with parallels to adipate
metabolism. Fast growth required the additional disruption
of the regulator-encoding psrA, which upregulates redundant
β-oxidation genes. This knowledge enabled the rational
reverse engineering of a strain that can not only use
adipate, but also other medium-chain-length dicarboxylates
like suberate and sebacate. The reverse engineered strain
grows on adipate with a rate of 0.35 ± 0.01 h−1,
reaching a final biomass yield of 0.27 ± 0.00 gCDW
gadipate−1. In a nitrogen-limited medium this strain
produced polyhydroxyalkanoates from adipate up to $25\%$ of
its CDW. This proves its applicability for the upcycling of
mixtures of polymers made from fossile resources into
biodegradable counterparts.},
cin = {IBG-1},
ddc = {610},
cid = {I:(DE-Juel1)IBG-1-20101118},
pnm = {2172 - Utilization of renewable carbon and energy sources
and engineering of ecosystem functions (POF4-217)},
pid = {G:(DE-HGF)POF4-2172},
typ = {PUB:(DE-HGF)16},
pubmed = {33965615},
UT = {WOS:000694909400004},
doi = {10.1016/j.ymben.2021.05.001},
url = {https://juser.fz-juelich.de/record/892434},
}