Laboratory evolution reveals the metabolic and regulatory basis of ethylene glycol metabolism by Pseudomonas putida KT2440

Li, Wing ‐Jin; Jayakody, Lahiru N.; Franden, Mary Ann; Wehrmann, Matthias; Daun, Tristan; Hauer, Bernhard; Blank, Lars M.; Beckham, Gregg T.; Klebensberger, Janosch; Wierckx, Nick

doi:10.1111/1462-2920.14703

Title: Laboratory evolution reveals the metabolic and regulatory basis of ethylene glycol metabolism by Pseudomonas putida KT2440

Journal Article · 05 June 2019 · Environmental Microbiology

DOI: https://doi.org/10.1111/1462-2920.14703 · OSTI ID:1526901

^[1]; Jayakody, Lahiru N. ^[2]; Franden, Mary Ann ^[2]; Wehrmann, Matthias ^[3]; Daun, Tristan ^[1]; Hauer, Bernhard ^[3]; Blank, Lars M. ^[1]; Beckham, Gregg T. ^[2]; Klebensberger, Janosch ^[3]; Wierckx, Nick ^[4]

RWTH Aachen Univ., Aachen (Germany)
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Univ. of Stuttgart, Stuttgart (Germany)
RWTH Aachen Univ., Aachen (Germany); Forschungszentrum Jülich, Jülich (Germany)

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, but 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.

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Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Energy Efficiency and Renewable Energy (EERE), Office of Sustainable Transportation and Fuels. Bioenergy Technologies Office (BETO); USDOE

Grant/Contract Number:: AC36-08GO28308; AC36‐08GO28308

OSTI ID:: 1526901

Alternate ID(s):: OSTI ID: 1542549

Report Number(s):: NREL/JA-2A00-74156

Journal Information:: Environmental Microbiology, Vol. 21; ISSN 1462-2912

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 62 works

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09 BIOMASS FUELS
59 BASIC BIOLOGICAL SCIENCES
Pseudomonas putida
ethylene glycol
glyoxylate
adaptive laboratory evolution
genomics
reverse engineering

Title: Laboratory evolution reveals the metabolic and regulatory basis of ethylene glycol metabolism by Pseudomonas putida KT2440

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