Publication Details

Category Text Publication
Reference Category Journals
DOI 10.1186/s12934-024-02509-8
Licence creative commons licence
Title (Primary) Systems biology of electrogenic Pseudomonas putida - multi-omics insights and metabolic engineering for enhanced 2-ketogluconate production
Author Weimer, A.; Pause, L.; Ries, F.; Kohlstedt, M.; Adrian, L.; Krömer, J.; Lai, B. ORCID logo ; Wittmann, C.
Source Titel Microbial Cell Factories
Year 2024
Department MIBITECH; MEB
Volume 23
Page From art. 246
Language englisch
Topic T7 Bioeconomy
Supplements https://static-content.springer.com/esm/art%3A10.1186%2Fs12934-024-02509-8/MediaObjects/12934_2024_2509_MOESM1_ESM.pdf
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Keywords Pseudomonas putida KT2440; Bio-electrochemical system; Systems biology; Electron transfer; Anode; 2-ketogluconate; Ribosome; Bio-production; Anoxic metabolism; Redox mediator
Abstract Background
Pseudomonas putida KT2440 has emerged as a promising host for industrial bioproduction. However, its strictly aerobic nature limits the scope of applications. Remarkably, this microbe exhibits high bioconversion efficiency when cultured in an anoxic bio-electrochemical system (BES), where the anode serves as the terminal electron acceptor instead of oxygen. This environment facilitates the synthesis of commercially attractive chemicals, including 2-ketogluconate (2KG). To better understand this interesting electrogenic phenotype, we studied the BES-cultured strain on a systems level through multi-omics analysis. Inspired by our findings, we constructed novel mutants aimed at improving 2KG production.

Results
When incubated on glucose, P. putida KT2440 did not grow but produced significant amounts of 2KG, along with minor amounts of gluconate, acetate, pyruvate, succinate, and lactate. 13C tracer studies demonstrated that these products are partially derived from biomass carbon, involving proteins and lipids. Over time, the cells exhibited global changes on both the transcriptomic and proteomic levels, including the shutdown of translation and cell motility, likely to conserve energy. These adaptations enabled the cells to maintain significant metabolic activity for several weeks. Acetate formation was shown to contribute to energy supply. Mutants deficient in acetate production demonstrated superior 2KG production in terms of titer, yield, and productivity. The ∆aldBI ∆aldBII double deletion mutant performed best, accumulating 2KG at twice the rate of the wild type and with an increased yield (0.96 mol/mol).

Conclusions
By integrating transcriptomic, proteomic, and metabolomic analyses, this work provides the first systems biology insight into the electrogenic phenotype of P. putida KT2440. Adaptation to anoxic-electrogenic conditions involved coordinated changes in energy metabolism, enabling cells to sustain metabolic activity for extended periods. The metabolically engineered mutants are promising for enhanced 2KG production under these conditions. The attenuation of acetate synthesis represents the first systems biology-informed metabolic engineering strategy for enhanced 2KG production in P. putida. This non-growth anoxic-electrogenic mode expands our understanding of the interplay between growth, glucose phosphorylation, and glucose oxidation into gluconate and 2KG in P. putida.
Persistent UFZ Identifier https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=29637
Weimer, A., Pause, L., Ries, F., Kohlstedt, M., Adrian, L., Krömer, J., Lai, B., Wittmann, C. (2024):
Systems biology of electrogenic Pseudomonas putida - multi-omics insights and metabolic engineering for enhanced 2-ketogluconate production
Microb. Cell. Fact. 23 , art. 246 10.1186/s12934-024-02509-8