Metabolic engineering of Pseudomonas putida for increased polyhydroxyalkanoate production from lignin
Abstract
Microbial conversion offers a promising strategy for overcoming the intrinsic heterogeneity of the plant biopolymer, lignin. Soil microbes that natively harbour aromatic-catabolic pathways are natural choices for chassis strains, and Pseudomonas putida KT2440 has emerged as a viable whole-cell biocatalyst for funnelling lignin-derived compounds to value-added products, including its native carbon storage product, medium-chain-length polyhydroxyalkanoates (mcl-PHA). In this work, a series of metabolic engineering targets to improve mcl-PHA production are combined in the P. putida chromosome and evaluated in strains growing in a model aromatic compound, p-coumaric acid, and in lignin streams. Specifically, the PHA depolymerase gene phaZ was knocked out, and the genes involved in ..beta..-oxidation (fadBA1 and fadBA2) were deleted. Additionally, to increase carbon flux into mcl-PHA biosynthesis, phaG, alkK, phaC1 and phaC2 were overexpressed. The best performing strain - which contains all the genetic modifications detailed above - demonstrated a 53% and 200% increase in mcl-PHA titre (g l-1) and a 20% and 100% increase in yield (g mcl-PHA per g cell dry weight) from p-coumaric acid and lignin, respectively, compared with the wild type strain. Overall, these results present a promising strain to be employed in further process development for enhancing mcl-PHA production from aromatic compoundsmore »
- Authors:
-
- National Renewable Energy Lab. (NREL), Golden, CO (United States). National Bioenergy Center
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division
- Publication Date:
- Research Org.:
- Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States); National Renewable Energy Laboratory (NREL), Golden, CO (United States)
- Sponsoring Org.:
- USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office
- OSTI Identifier:
- 1559676
- Alternate Identifier(s):
- OSTI ID: 1562862
- Report Number(s):
- NREL/JA-5100-74637
Journal ID: ISSN 1751-7915
- Grant/Contract Number:
- AC05-00OR22725; AC36‐08GO28308
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Microbial Biotechnology (Online)
- Additional Journal Information:
- Journal Name: Microbial Biotechnology (Online); Journal Volume: 13; Journal Issue: 1; Journal ID: ISSN 1751-7915
- Publisher:
- Wiley
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; metabolic engineering; Pseudomonas putida; microbial conversion; lignin; PHA; hydrocarbon fuels; 09 BIOMASS FUELS
Citation Formats
Salvachúa, Davinia, Rydzak, Thomas, Auwae, Raquel, De Capite, Annette, Black, Brenna A., Bouvier, Jason T., Cleveland, Nicholas S., Elmore, Joshua R., Huenemann, Jay D., Katahira, Rui, Michener, William E., Peterson, Darren J., Rohrer, Holly, Vardon, Derek R., Beckham, Gregg T., and Guss, Adam M. Metabolic engineering of Pseudomonas putida for increased polyhydroxyalkanoate production from lignin. United States: N. p., 2019.
Web. doi:10.1111/1751-7915.13481.
Salvachúa, Davinia, Rydzak, Thomas, Auwae, Raquel, De Capite, Annette, Black, Brenna A., Bouvier, Jason T., Cleveland, Nicholas S., Elmore, Joshua R., Huenemann, Jay D., Katahira, Rui, Michener, William E., Peterson, Darren J., Rohrer, Holly, Vardon, Derek R., Beckham, Gregg T., & Guss, Adam M. Metabolic engineering of Pseudomonas putida for increased polyhydroxyalkanoate production from lignin. United States. https://doi.org/10.1111/1751-7915.13481
Salvachúa, Davinia, Rydzak, Thomas, Auwae, Raquel, De Capite, Annette, Black, Brenna A., Bouvier, Jason T., Cleveland, Nicholas S., Elmore, Joshua R., Huenemann, Jay D., Katahira, Rui, Michener, William E., Peterson, Darren J., Rohrer, Holly, Vardon, Derek R., Beckham, Gregg T., and Guss, Adam M. Fri .
"Metabolic engineering of Pseudomonas putida for increased polyhydroxyalkanoate production from lignin". United States. https://doi.org/10.1111/1751-7915.13481. https://www.osti.gov/servlets/purl/1559676.
@article{osti_1559676,
title = {Metabolic engineering of Pseudomonas putida for increased polyhydroxyalkanoate production from lignin},
author = {Salvachúa, Davinia and Rydzak, Thomas and Auwae, Raquel and De Capite, Annette and Black, Brenna A. and Bouvier, Jason T. and Cleveland, Nicholas S. and Elmore, Joshua R. and Huenemann, Jay D. and Katahira, Rui and Michener, William E. and Peterson, Darren J. and Rohrer, Holly and Vardon, Derek R. and Beckham, Gregg T. and Guss, Adam M.},
abstractNote = {Microbial conversion offers a promising strategy for overcoming the intrinsic heterogeneity of the plant biopolymer, lignin. Soil microbes that natively harbour aromatic-catabolic pathways are natural choices for chassis strains, and Pseudomonas putida KT2440 has emerged as a viable whole-cell biocatalyst for funnelling lignin-derived compounds to value-added products, including its native carbon storage product, medium-chain-length polyhydroxyalkanoates (mcl-PHA). In this work, a series of metabolic engineering targets to improve mcl-PHA production are combined in the P. putida chromosome and evaluated in strains growing in a model aromatic compound, p-coumaric acid, and in lignin streams. Specifically, the PHA depolymerase gene phaZ was knocked out, and the genes involved in ..beta..-oxidation (fadBA1 and fadBA2) were deleted. Additionally, to increase carbon flux into mcl-PHA biosynthesis, phaG, alkK, phaC1 and phaC2 were overexpressed. The best performing strain - which contains all the genetic modifications detailed above - demonstrated a 53% and 200% increase in mcl-PHA titre (g l-1) and a 20% and 100% increase in yield (g mcl-PHA per g cell dry weight) from p-coumaric acid and lignin, respectively, compared with the wild type strain. Overall, these results present a promising strain to be employed in further process development for enhancing mcl-PHA production from aromatic compounds and lignin.},
doi = {10.1111/1751-7915.13481},
journal = {Microbial Biotechnology (Online)},
number = 1,
volume = 13,
place = {United States},
year = {Fri Aug 30 00:00:00 EDT 2019},
month = {Fri Aug 30 00:00:00 EDT 2019}
}
Web of Science
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Works referencing / citing this record:
Development of a CRISPR/Cas9n-based tool for metabolic engineering of Pseudomonas putida for ferulic acid-to-polyhydroxyalkanoate bioconversion
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