Prediction of reaction knockouts to maximize succinate production by Actinobacillus succinogenes
Abstract
Succinate is a precursor of multiple commodity chemicals and bio-based succinate production is an active area of industrial bioengineering research. One of the most important microbial strains for bio-based production of succinate is the capnophilic gram-negative bacterium Actinobacillus succinogenes, which naturally produces succinate by a mixed-acid fermentative pathway. To engineer A. succinogenes to improve succinate yields during mixed acid fermentation, it is important to have a detailed understanding of the metabolic flux distribution in A. succinogenes when grown in suitable media. To this end, we have developed a detailed stoichiometric model of the A. succinogenes central metabolism that includes the biosynthetic pathways for the main components of biomass - namely glycogen, amino acids, DNA, RNA, lipids and UDP-N-Acetyl-a-D-glucosamine. We have validated our model by comparing model predictions generated via flux balance analysis with experimental results on mixed acid fermentation. Moreover, we have used the model to predict single and double reaction knockouts to maximize succinate production while maintaining growth viability. According to our model, succinate production can be maximized by knocking out either of the reactions catalyzed by the PTA (phosphate acetyltransferase) and ACK (acetyl kinase) enzymes, whereas the double knockouts of PEPCK (phosphoenolpyruvate carboxykinase) and PTA or PEPCK andmore »
- Authors:
- Publication Date:
- Research Org.:
- 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 (BETO)
- OSTI Identifier:
- 1418730
- Alternate Identifier(s):
- OSTI ID: 1423185
- Report Number(s):
- NREL/JA-2700-70444
Journal ID: ISSN 1932-6203; 10.1371/journal.pone.0189144
- Grant/Contract Number:
- AC36-08GO28308
- Resource Type:
- Published Article
- Journal Name:
- PLoS ONE
- Additional Journal Information:
- Journal Name: PLoS ONE Journal Volume: 13 Journal Issue: 1; Journal ID: ISSN 1932-6203
- Publisher:
- Public Library of Science (PLoS)
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; bicarbonates; fermentation; metabolic networks; glucose; enzymes; ethanol; glycogens; metabolic pathways
Citation Formats
Nag, Ambarish, St. John, Peter C., Crowley, Michael F., Bomble, Yannick J., and Du, ed., Chenyu. Prediction of reaction knockouts to maximize succinate production by Actinobacillus succinogenes. United States: N. p., 2018.
Web. doi:10.1371/journal.pone.0189144.
Nag, Ambarish, St. John, Peter C., Crowley, Michael F., Bomble, Yannick J., & Du, ed., Chenyu. Prediction of reaction knockouts to maximize succinate production by Actinobacillus succinogenes. United States. https://doi.org/10.1371/journal.pone.0189144
Nag, Ambarish, St. John, Peter C., Crowley, Michael F., Bomble, Yannick J., and Du, ed., Chenyu. Tue .
"Prediction of reaction knockouts to maximize succinate production by Actinobacillus succinogenes". United States. https://doi.org/10.1371/journal.pone.0189144.
@article{osti_1418730,
title = {Prediction of reaction knockouts to maximize succinate production by Actinobacillus succinogenes},
author = {Nag, Ambarish and St. John, Peter C. and Crowley, Michael F. and Bomble, Yannick J. and Du, ed., Chenyu},
abstractNote = {Succinate is a precursor of multiple commodity chemicals and bio-based succinate production is an active area of industrial bioengineering research. One of the most important microbial strains for bio-based production of succinate is the capnophilic gram-negative bacterium Actinobacillus succinogenes, which naturally produces succinate by a mixed-acid fermentative pathway. To engineer A. succinogenes to improve succinate yields during mixed acid fermentation, it is important to have a detailed understanding of the metabolic flux distribution in A. succinogenes when grown in suitable media. To this end, we have developed a detailed stoichiometric model of the A. succinogenes central metabolism that includes the biosynthetic pathways for the main components of biomass - namely glycogen, amino acids, DNA, RNA, lipids and UDP-N-Acetyl-a-D-glucosamine. We have validated our model by comparing model predictions generated via flux balance analysis with experimental results on mixed acid fermentation. Moreover, we have used the model to predict single and double reaction knockouts to maximize succinate production while maintaining growth viability. According to our model, succinate production can be maximized by knocking out either of the reactions catalyzed by the PTA (phosphate acetyltransferase) and ACK (acetyl kinase) enzymes, whereas the double knockouts of PEPCK (phosphoenolpyruvate carboxykinase) and PTA or PEPCK and ACK enzymes are the most effective in increasing succinate production.},
doi = {10.1371/journal.pone.0189144},
journal = {PLoS ONE},
number = 1,
volume = 13,
place = {United States},
year = {Tue Jan 30 00:00:00 EST 2018},
month = {Tue Jan 30 00:00:00 EST 2018}
}
https://doi.org/10.1371/journal.pone.0189144
Web of Science
Works referenced in this record:
A protocol for generating a high-quality genome-scale metabolic reconstruction
journal, January 2010
- Thiele, Ines; Palsson, Bernhard Ø
- Nature Protocols, Vol. 5, Issue 1
Succinic acid production with Actinobacillus succinogenes: rate and yield analysis of chemostat and biofilm cultures
journal, August 2014
- Brink, Hendrik Gideon; Nicol, Willie
- Microbial Cell Factories, Vol. 13, Issue 1
Development of a Markerless Knockout Method for Actinobacillus succinogenes
journal, March 2014
- Joshi, Rajasi V.; Schindler, Bryan D.; McPherson, Nikolas R.
- Applied and Environmental Microbiology, Vol. 80, Issue 10
Environmental and physiological factors affecting the succinate product ratio during carbohydrate fermentation by Actinobacillus sp. 130Z
journal, May 1997
- der Werf, M. J. Van; Guettler, Michael V.; Jain, Mahendra K.
- Archives of Microbiology, Vol. 167, Issue 6
Flux balance analysis in the era of metabolomics
journal, March 2006
- Lee, J. M.
- Briefings in Bioinformatics, Vol. 7, Issue 2
OpenFLUX: efficient modelling software for 13C-based metabolic flux analysis
journal, January 2009
- Quek, Lake-Ee; Wittmann, Christoph; Nielsen, Lars K.
- Microbial Cell Factories, Vol. 8, Issue 1
Insights into Actinobacillus succinogenes Fermentative Metabolism in a Chemically Defined Growth Medium
journal, November 2005
- McKinlay, J. B.; Zeikus, J. G.; Vieille, C.
- Applied and Environmental Microbiology, Vol. 71, Issue 11
Determining Actinobacillus succinogenes metabolic pathways and fluxes by NMR and GC-MS analyses of 13C-labeled metabolic product isotopomers
journal, March 2007
- McKinlay, James B.; Shachar-Hill, Yair; Zeikus, J. Gregory
- Metabolic Engineering, Vol. 9, Issue 2
13C-metabolic flux analysis of Actinobacillus succinogenes fermentative metabolism at different NaHCO3 and H2 concentrations
journal, January 2008
- McKinlay, James B.; Vieille, Claire
- Metabolic Engineering, Vol. 10, Issue 1, p. 55-68
OpenMebius: An Open Source Software for Isotopically Nonstationary 13 C-Based Metabolic Flux Analysis
journal, January 2014
- Kajihata, Shuichi; Furusawa, Chikara; Matsuda, Fumio
- BioMed Research International, Vol. 2014
Metabolic networks in motion: 13 C‐based flux analysis
journal, January 2006
- Sauer, Uwe
- Molecular Systems Biology, Vol. 2, Issue 1
Reconstructing the metabolic network of a bacterium from its genome
journal, November 2005
- Francke, Christof; Siezen, Roland J.; Teusink, Bas
- Trends in Microbiology, Vol. 13, Issue 11
Diversity of flux distribution in central carbon metabolism of S. cerevisiae strains from diverse environments
journal, April 2016
- Nidelet, Thibault; Brial, Pascale; Camarasa, Carole
- Microbial Cell Factories, Vol. 15, Issue 1
Quantitative prediction of cellular metabolism with constraint-based models: the COBRA Toolbox v2.0
journal, August 2011
- Schellenberger, Jan; Que, Richard; Fleming, Ronan M. T.
- Nature Protocols, Vol. 6, Issue 9
The effects of alternate optimal solutions in constraint-based genome-scale metabolic models
journal, October 2003
- Mahadevan, R.; Schilling, C. H.
- Metabolic Engineering, Vol. 5, Issue 4
Stoichiometric modelling of cell metabolism
journal, January 2008
- Llaneras, Francisco; Picó, Jesús
- Journal of Bioscience and Bioengineering, Vol. 105, Issue 1
Systems metabolic engineering: Genome-scale models and beyond
journal, February 2010
- Blazeck, John; Alper, Hal
- Biotechnology Journal, Vol. 5, Issue 7
Comparison of network-based pathway analysis methods
journal, August 2004
- Papin, Jason A.; Stelling, Joerg; Price, Nathan D.
- Trends in Biotechnology, Vol. 22, Issue 8
The Metabolic Network of Synechocystis sp. PCC 6803: Systemic Properties of Autotrophic Growth
journal, July 2010
- Knoop, Henning; Zilliges, Yvonne; Lockau, Wolfgang
- Plant Physiology, Vol. 154, Issue 1
What is flux balance analysis?
journal, March 2010
- Orth, Jeffrey D.; Thiele, Ines; Palsson, Bernhard Ø
- Nature Biotechnology, Vol. 28, Issue 3
Constraint-based models predict metabolic and associated cellular functions
journal, January 2014
- Bordbar, Aarash; Monk, Jonathan M.; King, Zachary A.
- Nature Reviews Genetics, Vol. 15, Issue 2
Omic data from evolved E. coli are consistent with computed optimal growth from genome‐scale models
journal, January 2010
- Lewis, Nathan E.; Hixson, Kim K.; Conrad, Tom M.
- Molecular Systems Biology, Vol. 6, Issue 1
A genomic perspective on the potential of Actinobacillus succinogenes for industrial succinate production
journal, January 2010
- McKinlay, James B.; Laivenieks, Maris; Schindler, Bryan D.
- BMC Genomics, Vol. 11, Issue 1
Actinobacillus succinogenes sp. nov., a novel succinic-acid-producing strain from the bovine rumen
journal, January 1999
- Guettler, M. V.; Rumler, D.; Jain, M. K.
- International Journal of Systematic Bacteriology, Vol. 49, Issue 1
Computational tools for metabolic engineering
journal, May 2012
- Copeland, Wilbert B.; Bartley, Bryan A.; Chandran, Deepak
- Metabolic Engineering, Vol. 14, Issue 3
Metabolic capabilities of Actinobacillus succinogenes for succinic acid production
journal, December 2014
- Rafieenia, R.
- Brazilian Journal of Chemical Engineering, Vol. 31, Issue 4
Production of succinic acid by bacterial fermentation
journal, July 2006
- Song, Hyohak; Lee, Sang Yup
- Enzyme and Microbial Technology, Vol. 39, Issue 3
COBRApy: COnstraints-Based Reconstruction and Analysis for Python
journal, January 2013
- Ebrahim, Ali; Lerman, Joshua A.; Palsson, Bernhard O.
- BMC Systems Biology, Vol. 7, Issue 1
Saccharomyces cerevisiae phenotypes can be predicted by using constraint-based analysis of a genome-scale reconstructed metabolic network
journal, October 2003
- Famili, I.; Forster, J.; Nielsen, J.
- Proceedings of the National Academy of Sciences, Vol. 100, Issue 23