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Title: 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 » and lignin.« less

Authors:
ORCiD logo [1];  [2];  [2];  [2];  [1];  [2]; ORCiD logo [1];  [2];  [2];  [1];  [1];  [1];  [1]; ORCiD logo [1]; ORCiD logo [1];  [2]
  1. National Renewable Energy Laboratory (NREL), Golden, CO (United States)
  2. Oak Ridge National Laboratory
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Energy Efficiency and Renewable Energy (EERE), Bioenergy Technologies Office (EE-3B)
OSTI Identifier:
1562862
Report Number(s):
NREL/JA-5100-74637
DOE Contract Number:  
AC36-08GO28308
Resource Type:
Journal Article
Journal Name:
Microbial Biotechnology
Additional Journal Information:
Journal Name: Microbial Biotechnology
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; metabolic engineering; Pseudomonas putida; microbial conversion; lignin; PHA; hydrocarbon fuels

Citation Formats

Salvachua Rodriguez, Davinia, Rydzak, Thomas, Auwae, Raquel, De Capite, Annette, Pleitner, Brenna P, Bouvier, Jason T., Cleveland, Nicholas S, Elmore, Joshua R., Huenemann, Jay D., Katahira, Rui, Michener, William E, Peterson, Darren J, Rohrer, Holly K, 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.
Salvachua Rodriguez, Davinia, Rydzak, Thomas, Auwae, Raquel, De Capite, Annette, Pleitner, Brenna P, Bouvier, Jason T., Cleveland, Nicholas S, Elmore, Joshua R., Huenemann, Jay D., Katahira, Rui, Michener, William E, Peterson, Darren J, Rohrer, Holly K, Vardon, Derek R, Beckham, Gregg T, & Guss, Adam M. Metabolic Engineering of Pseudomonas putida for Increased Polyhydroxyalkanoate Production from Lignin. United States. doi:10.1111/1751-7915.13481.
Salvachua Rodriguez, Davinia, Rydzak, Thomas, Auwae, Raquel, De Capite, Annette, Pleitner, Brenna P, Bouvier, Jason T., Cleveland, Nicholas S, Elmore, Joshua R., Huenemann, Jay D., Katahira, Rui, Michener, William E, Peterson, Darren J, Rohrer, Holly K, Vardon, Derek R, Beckham, Gregg T, and Guss, Adam M. Fri . "Metabolic Engineering of Pseudomonas putida for Increased Polyhydroxyalkanoate Production from Lignin". United States. doi:10.1111/1751-7915.13481.
@article{osti_1562862,
title = {Metabolic Engineering of Pseudomonas putida for Increased Polyhydroxyalkanoate Production from Lignin},
author = {Salvachua Rodriguez, Davinia and Rydzak, Thomas and Auwae, Raquel and De Capite, Annette and Pleitner, Brenna P 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 K 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},
number = ,
volume = ,
place = {United States},
year = {2019},
month = {8}
}

Works referenced in this record:

The tac promoter: a functional hybrid derived from the trp and lac promoters
journal, January 1983

  • de Boer, H. A.; Comstock, L. J.; Vasser, M.
  • Proceedings of the National Academy of Sciences, Vol. 80, Issue 1, p. 21-25
  • DOI: 10.1073/pnas.80.1.21

Lignin valorization through integrated biological funneling and chemical catalysis
journal, August 2014

  • Linger, J. G.; Vardon, D. R.; Guarnieri, M. T.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 33, p. 12013-12018
  • DOI: 10.1073/pnas.1410657111

Development of a broad-host-range sacB-based vector for unmarked allelic exchange
journal, January 2008


Lignin Valorization: Improving Lignin Processing in the Biorefinery
journal, May 2014

  • Ragauskas, A. J.; Beckham, G. T.; Biddy, M. J.
  • Science, Vol. 344, Issue 6185, p. 1246843-1246843
  • DOI: 10.1126/science.1246843

Development of a New Strategy for Production of Medium-Chain-Length Polyhydroxyalkanoates by Recombinant Escherichia coli via Inexpensive Non-Fatty Acid Feedstocks
journal, November 2011

  • Wang, Qin; Tappel, Ryan C.; Zhu, Chengjun
  • Applied and Environmental Microbiology, Vol. 78, Issue 2, p. 519-527
  • DOI: 10.1128/AEM.07020-11