TY  - JOUR
AU  - Ackermann, Yannic S.
AU  - Li, Wing-Jin
AU  - Op de Hipt, Leonie
AU  - Niehoff, Paul-Joachim
AU  - Casey, William
AU  - Polen, Tino
AU  - Köbbing, Sebastian
AU  - Ballerstedt, Hendrik
AU  - Wynands, Benedikt
AU  - O'Connor, Kevin
AU  - Blank, Lars M.
AU  - Wierckx, Nick
TI  - Engineering adipic acid metabolism in Pseudomonas putida
JO  - Metabolic engineering
VL  - 67
SN  - 1096-7176
CY  - Orlando, Fla.
PB  - Academic Press
M1  - FZJ-2021-02080
SP  - 29-40
PY  - 2021
N1  - Biotechnologie 1
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - 33965615
UR  - <Go to ISI:>//WOS:000694909400004
DO  - DOI:10.1016/j.ymben.2021.05.001
UR  - https://juser.fz-juelich.de/record/892434
ER  -