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Synthetically-primed adaptation of Pseudomonas putida to a non-native substrate D-xylose.

Authors :
Dvořák, Pavel
Burýšková, Barbora
Popelářová, Barbora
Ebert, Birgitta E.
Botka, Tibor
Bujdoš, Dalimil
Sánchez-Pascuala, Alberto
Schöttler, Hannah
Hayen, Heiko
de Lorenzo, Víctor
Blank, Lars M.
Benešík, Martin
Source :
Nature Communications; 3/26/2024, Vol. 15 Issue 1, p1-18, 18p
Publication Year :
2024

Abstract

To broaden the substrate scope of microbial cell factories towards renewable substrates, rational genetic interventions are often combined with adaptive laboratory evolution (ALE). However, comprehensive studies enabling a holistic understanding of adaptation processes primed by rational metabolic engineering remain scarce. The industrial workhorse Pseudomonas putida was engineered to utilize the non-native sugar D-xylose, but its assimilation into the bacterial biochemical network via the exogenous xylose isomerase pathway remained unresolved. Here, we elucidate the xylose metabolism and establish a foundation for further engineering followed by ALE. First, native glycolysis is derepressed by deleting the local transcriptional regulator gene hexR. We then enhance the pentose phosphate pathway by implanting exogenous transketolase and transaldolase into two lag-shortened strains and allow ALE to finetune the rewired metabolism. Subsequent multilevel analysis and reverse engineering provide detailed insights into the parallel paths of bacterial adaptation to the non-native carbon source, highlighting the enhanced expression of transaldolase and xylose isomerase along with derepressed glycolysis as key events during the process. Pseudomonas putida is becoming a host of choice for the valorization of lignocellulosic substrates. Here, the authors provide insight into the adaptation of this bacterium to the non-native substrate D-xylose, enabled by metabolic engineering and adaptive laboratory evolution. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
20411723
Volume :
15
Issue :
1
Database :
Complementary Index
Journal :
Nature Communications
Publication Type :
Academic Journal
Accession number :
176264915
Full Text :
https://doi.org/10.1038/s41467-024-46812-9