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Title: Methyl ketone production by Pseudomonas putida is enhanced by plant-derived amino acids

Abstract

Plants are an attractive source of renewable carbon for conversion to biofuels and bio-based chemicals. Conversion strategies often use a fraction of the biomass, focusing on sugars from cellulose and hemicellulose. Strategies that use plant components, such as aromatics and amino acids, may improve the efficiency of biomass conversion. Pseudomonas putida is a promising host for its ability to metabolize a wide variety of organic compounds. P. putida was engineered to produce methyl ketones, which are promising diesel blendstocks and potential platform chemicals, from glucose and lignin-related aromatics. Unexpectedly, P. putida methyl ketone production using Arabidopsis thaliana hydrolysates was enhanced 2-5-fold compared with sugar controls derived from engineered plants that overproduce lignin-related aromatics. This enhancement was more pronounced (~seven-fold increase) with hydrolysates from nonengineered switchgrass. Proteomic analysis of the methyl ketone-producing P. putida suggested that plant-derived amino acids may be the source of this enhancement. Mass spectrometry-based measurements of plant-derived amino acids demonstrated a high correlation between methyl ketone production and amino acid concentration in plant hydrolysates. Amendment of glucose-containing minimal media with a defined mixture of amino acids similar to those found in the hydrolysates studied led to a nine-fold increase in methyl ketone titer (1.1 g/L).

Authors:
 [1];  [1];  [1];  [1];  [1]; ORCiD logo [2];  [3];  [3];  [4];  [5];  [1]; ORCiD logo [1]
  1. Joint BioEnergy InstituteEmeryville California; Biological Systems and Engineering DivisionLawrence Berkeley National LaboratoryBerkeley California
  2. Joint BioEnergy InstituteEmeryville California; Earth and Environmental SciencesLawrence Berkeley National LaboratoryBerkeley California
  3. Joint BioEnergy InstituteEmeryville California; Environmental Genomics and Systems Biology DivisionLawrence Berkeley National LaboratoryBerkeley California
  4. Joint BioEnergy InstituteEmeryville California; Molecular Biophysics and Integrated Bioimaging DivisionLawrence Berkeley National LaboratoryBerkeley California
  5. Joint BioEnergy InstituteEmeryville California; Biological Systems and Engineering DivisionLawrence Berkeley National LaboratoryBerkeley California; Environmental Genomics and Systems Biology DivisionLawrence Berkeley National LaboratoryBerkeley California
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1545149
DOE Contract Number:  
AC02-05CH11231
Resource Type:
Journal Article
Journal Name:
Biotechnology and Bioengineering
Additional Journal Information:
Journal Volume: 116; Journal Issue: 8; Journal ID: ISSN 0006-3592
Publisher:
Wiley
Country of Publication:
United States
Language:
English

Citation Formats

Dong, Jie, Chen, Yan, Benites, Veronica Teixeira, Baidoo, Edward E. K., Petzold, Christopher J., Beller, Harry R., Eudes, Aymerick, Scheller, Henrik V., Adams, Paul D., Mukhopadhyay, Aindrila, Simmons, Blake A., and Singer, Steven W. Methyl ketone production by Pseudomonas putida is enhanced by plant-derived amino acids. United States: N. p., 2019. Web. doi:10.1002/bit.26995.
Dong, Jie, Chen, Yan, Benites, Veronica Teixeira, Baidoo, Edward E. K., Petzold, Christopher J., Beller, Harry R., Eudes, Aymerick, Scheller, Henrik V., Adams, Paul D., Mukhopadhyay, Aindrila, Simmons, Blake A., & Singer, Steven W. Methyl ketone production by Pseudomonas putida is enhanced by plant-derived amino acids. United States. doi:10.1002/bit.26995.
Dong, Jie, Chen, Yan, Benites, Veronica Teixeira, Baidoo, Edward E. K., Petzold, Christopher J., Beller, Harry R., Eudes, Aymerick, Scheller, Henrik V., Adams, Paul D., Mukhopadhyay, Aindrila, Simmons, Blake A., and Singer, Steven W. Mon . "Methyl ketone production by Pseudomonas putida is enhanced by plant-derived amino acids". United States. doi:10.1002/bit.26995.
@article{osti_1545149,
title = {Methyl ketone production by Pseudomonas putida is enhanced by plant-derived amino acids},
author = {Dong, Jie and Chen, Yan and Benites, Veronica Teixeira and Baidoo, Edward E. K. and Petzold, Christopher J. and Beller, Harry R. and Eudes, Aymerick and Scheller, Henrik V. and Adams, Paul D. and Mukhopadhyay, Aindrila and Simmons, Blake A. and Singer, Steven W.},
abstractNote = {Plants are an attractive source of renewable carbon for conversion to biofuels and bio-based chemicals. Conversion strategies often use a fraction of the biomass, focusing on sugars from cellulose and hemicellulose. Strategies that use plant components, such as aromatics and amino acids, may improve the efficiency of biomass conversion. Pseudomonas putida is a promising host for its ability to metabolize a wide variety of organic compounds. P. putida was engineered to produce methyl ketones, which are promising diesel blendstocks and potential platform chemicals, from glucose and lignin-related aromatics. Unexpectedly, P. putida methyl ketone production using Arabidopsis thaliana hydrolysates was enhanced 2-5-fold compared with sugar controls derived from engineered plants that overproduce lignin-related aromatics. This enhancement was more pronounced (~seven-fold increase) with hydrolysates from nonengineered switchgrass. Proteomic analysis of the methyl ketone-producing P. putida suggested that plant-derived amino acids may be the source of this enhancement. Mass spectrometry-based measurements of plant-derived amino acids demonstrated a high correlation between methyl ketone production and amino acid concentration in plant hydrolysates. Amendment of glucose-containing minimal media with a defined mixture of amino acids similar to those found in the hydrolysates studied led to a nine-fold increase in methyl ketone titer (1.1 g/L).},
doi = {10.1002/bit.26995},
journal = {Biotechnology and Bioengineering},
issn = {0006-3592},
number = 8,
volume = 116,
place = {United States},
year = {2019},
month = {4}
}

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