Laboratory evolution reveals the metabolic and regulatory basis of ethylene glycol metabolism by Pseudomonas putida KT2440
| Autor: | Bernhard Hauer, Mary Ann Franden, Lahiru N. Jayakody, Lars M. Blank, Gregg T. Beckham, Tristan Daun, Wing-Jin Li, Janosch Klebensberger, Nick Wierckx, Matthias Wehrmann |
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| Rok vydání: | 2019 |
| Předmět: |
Purine
Ethylene Glycol 0303 health sciences biology Pseudomonas putida 030306 microbiology Mutant Microbial metabolism Glyoxylate cycle Context (language use) Metabolism biology.organism_classification Microbiology Carbon 03 medical and health sciences chemistry.chemical_compound chemistry Biochemistry ddc:570 Environmental Pollutants Directed Molecular Evolution Ethylene glycol Ecology Evolution Behavior and Systematics 030304 developmental biology |
| Zdroj: | Environmental microbiology 21(10), 3669-3682 (2019). doi:10.1111/1462-2920.14703 |
| ISSN: | 1462-2920 1462-2912 |
| DOI: | 10.1111/1462-2920.14703 |
| Popis: | Pollution from ethylene glycol, and plastics containing this monomer, represent a significant environmental problem. The investigation of its microbial metabolism therefore provides insights into the environmental fate of this pollutant and also enables its utilization as a carbon source for microbial biotechnology. Here, we reveal the genomic and metabolic basis of ethylene glycol metabolism in Pseudomonas putida KT2440. Although this strain cannot grow on ethylene glycol as sole carbon source, it can be used to generate growth-enhancing reducing equivalents upon co-feeding with acetate. Mutants that utilize ethylene glycol as sole carbon source were isolated through adaptive laboratory evolution. Genomic analysis of these mutants revealed a central role of the transcriptional regulator GclR, which represses the glyoxylate carboligase pathway as part of a larger metabolic context of purine and allantoin metabolism. Secondary mutations in a transcriptional regulator encoded by PP_2046 and a porin encoded by PP_2662 further improved growth on ethylene glycol in evolved strains, likely by balancing fluxes through the initial oxidations of ethylene glycol to glyoxylate. With this knowledge, we reverse engineered an ethylene glycol utilizing strain and thus revealed the metabolic and regulatory basis that are essential for efficient ethylene glycol metabolism in P. putida KT2440. |
| Databáze: | OpenAIRE |
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