Development of a core Clostridium thermocellum kinetic metabolic model consistent with multiple genetic perturbations
Abstract
Background. Clostridium thermocellum is a Gram-positive anaerobe with the ability to hydrolyze and metabolize cellulose into biofuels such as ethanol, making it an attractive candidate for consolidated bioprocessing (CBP). At present, metabolic engineering in C. thermocellum is hindered due to the incomplete description of its metabolic repertoire and regulation within a predictive metabolic model. Genome-scale metabolic (GSM) models augmented with kinetic models of metabolism have been shown to be effective at recapitulating perturbed metabolic phenotypes. Results. In this effort, we first update a second-generation genome-scale metabolic model (iCth446) for C. thermocellum by correcting cofactor dependencies, restoring elemental and charge balances, and updating GAM and NGAM values to improve phenotype predictions. The iCth446 model is next used as a scaffold to develop a core kinetic model (k-ctherm118) of the C. thermocellum central metabolism using the Ensemble Modeling (EM) paradigm. Model parameterization is carried out by simultaneously imposing fermentation yield data in lactate, malate, acetate, and hydrogen production pathways for 19 measured metabolites spanning a library of 19 distinct single and multiple gene knockout mutants along with 18 intracellular metabolite concentration data for a Δgldh mutant and ten experimentally measured Michaelis–Menten kinetic parameters. Conclusions. The k-ctherm118 model captures significant metabolic changes causedmore »
- Authors:
- Publication Date:
- Research Org.:
- Pennsylvania State Univ., University Park, PA (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1618677
- Alternate Identifier(s):
- OSTI ID: 1393548
- Grant/Contract Number:
- SC0012377
- Resource Type:
- Published Article
- Journal Name:
- Biotechnology for Biofuels
- Additional Journal Information:
- Journal Name: Biotechnology for Biofuels Journal Volume: 10 Journal Issue: 1; Journal ID: ISSN 1754-6834
- Publisher:
- Springer Science + Business Media
- Country of Publication:
- Netherlands
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; Clostridium thermocellum; Genome-scale metabolic model; Kinetic model; Ensemble modeling; Nitrogen limitation; Ethanol stress
Citation Formats
Dash, Satyakam, Khodayari, Ali, Zhou, Jilai, Holwerda, Evert K., Olson, Daniel G., Lynd, Lee R., and Maranas, Costas D. Development of a core Clostridium thermocellum kinetic metabolic model consistent with multiple genetic perturbations. Netherlands: N. p., 2017.
Web. doi:10.1186/s13068-017-0792-2.
Dash, Satyakam, Khodayari, Ali, Zhou, Jilai, Holwerda, Evert K., Olson, Daniel G., Lynd, Lee R., & Maranas, Costas D. Development of a core Clostridium thermocellum kinetic metabolic model consistent with multiple genetic perturbations. Netherlands. https://doi.org/10.1186/s13068-017-0792-2
Dash, Satyakam, Khodayari, Ali, Zhou, Jilai, Holwerda, Evert K., Olson, Daniel G., Lynd, Lee R., and Maranas, Costas D. Tue .
"Development of a core Clostridium thermocellum kinetic metabolic model consistent with multiple genetic perturbations". Netherlands. https://doi.org/10.1186/s13068-017-0792-2.
@article{osti_1618677,
title = {Development of a core Clostridium thermocellum kinetic metabolic model consistent with multiple genetic perturbations},
author = {Dash, Satyakam and Khodayari, Ali and Zhou, Jilai and Holwerda, Evert K. and Olson, Daniel G. and Lynd, Lee R. and Maranas, Costas D.},
abstractNote = {Background. Clostridium thermocellum is a Gram-positive anaerobe with the ability to hydrolyze and metabolize cellulose into biofuels such as ethanol, making it an attractive candidate for consolidated bioprocessing (CBP). At present, metabolic engineering in C. thermocellum is hindered due to the incomplete description of its metabolic repertoire and regulation within a predictive metabolic model. Genome-scale metabolic (GSM) models augmented with kinetic models of metabolism have been shown to be effective at recapitulating perturbed metabolic phenotypes. Results. In this effort, we first update a second-generation genome-scale metabolic model (iCth446) for C. thermocellum by correcting cofactor dependencies, restoring elemental and charge balances, and updating GAM and NGAM values to improve phenotype predictions. The iCth446 model is next used as a scaffold to develop a core kinetic model (k-ctherm118) of the C. thermocellum central metabolism using the Ensemble Modeling (EM) paradigm. Model parameterization is carried out by simultaneously imposing fermentation yield data in lactate, malate, acetate, and hydrogen production pathways for 19 measured metabolites spanning a library of 19 distinct single and multiple gene knockout mutants along with 18 intracellular metabolite concentration data for a Δgldh mutant and ten experimentally measured Michaelis–Menten kinetic parameters. Conclusions. The k-ctherm118 model captures significant metabolic changes caused by (1) nitrogen limitation leading to increased yields for lactate, pyruvate, and amino acids, and (2) ethanol stress causing an increase in intracellular sugar phosphate concentrations (~1.5-fold) due to upregulation of cofactor pools. Robustness analysis of k-ctherm118 alludes to the presence of a secondary activity of ketol-acid reductoisomerase and possible regulation by valine and/or leucine pool levels. In addition, cross-validation and robustness analysis allude to missing elements in k-ctherm118 and suggest additional experiments to improve kinetic model prediction fidelity. Overall, the study quantitatively assesses the advantages of EM-based kinetic modeling towards improved prediction of C. thermocellum metabolism and develops a predictive kinetic model which can be used to design biofuel-overproducing strains.},
doi = {10.1186/s13068-017-0792-2},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 10,
place = {Netherlands},
year = {Tue May 02 00:00:00 EDT 2017},
month = {Tue May 02 00:00:00 EDT 2017}
}
https://doi.org/10.1186/s13068-017-0792-2
Web of Science
Works referenced in this record:
SMET: Systematic multiple enzyme targeting - a method to rationally design optimal strains for target chemical overproduction
journal, April 2013
- Flowers, David; Thompson, R. Adam; Birdwell, Douglas
- Biotechnology Journal, Vol. 8, Issue 5
Glycolysis without pyruvate kinase in Clostridium thermocellum
journal, January 2017
- Olson, Daniel G.; Hörl, Manuel; Fuhrer, Tobias
- Metabolic Engineering, Vol. 39
Amino acid transport by membrane vesicles of an obligate anaerobic bacterium, Clostridium acetobutylicum.
journal, February 1988
- Driessen, A. J.; Ubbink-Kok, T.; Konings, W. N.
- Journal of Bacteriology, Vol. 170, Issue 2
Metabolomics in systems microbiology
journal, February 2011
- Reaves, Marshall Louis; Rabinowitz, Joshua D.
- Current Opinion in Biotechnology, Vol. 22, Issue 1
Proteomic analysis of Clostridium thermocellum core metabolism: relative protein expression profiles and growth phase-dependent changes in protein expression
journal, January 2012
- Rydzak, Thomas; McQueen, Peter D.; Krokhin, Oleg V.
- BMC Microbiology, Vol. 12, Issue 1
Exploring complex cellular phenotypes and model-guided strain design with a novel genome-scale metabolic model of Clostridium thermocellum DSM 1313 implementing an adjustable cellulosome
journal, September 2016
- Thompson, R. Adam; Dahal, Sanjeev; Garcia, Sergio
- Biotechnology for Biofuels, Vol. 9, Issue 1
Clostridium thermocellum ATCC27405 transcriptomic, metabolomic and proteomic profiles after ethanol stress
journal, January 2012
- Yang, Shihui; Giannone, Richard J.; Dice, Lezlee
- BMC Genomics, Vol. 13, Issue 1
Metabolic selectivity and growth of Clostridium thermocellum in continuous culture under elevated hydrostatic pressure
journal, January 2004
- Bothun, G. D.; Knutson, B. L.; Berberich, J. A.
- Applied Microbiology and Biotechnology, Vol. 65, Issue 2
Elucidating central metabolic redox obstacles hindering ethanol production in Clostridium thermocellum
journal, November 2015
- Thompson, R. Adam; Layton, Donovan S.; Guss, Adam M.
- Metabolic Engineering, Vol. 32
CO 2 -fixing one-carbon metabolism in a cellulose-degrading bacterium Clostridium thermocellum
journal, October 2016
- Xiong, Wei; Lin, Paul P.; Magnusson, Lauren
- Proceedings of the National Academy of Sciences, Vol. 113, Issue 46
Cellulase, Clostridia, and Ethanol
journal, March 2005
- Demain, A. L.; Newcomb, M.; Wu, J. H. D.
- Microbiology and Molecular Biology Reviews, Vol. 69, Issue 1, p. 124-154
Characterization of Clostridium thermocellum strains with disrupted fermentation end-product pathways
journal, May 2013
- van der Veen, Douwe; Lo, Jonathan; Brown, Steven D.
- Journal of Industrial Microbiology & Biotechnology, Vol. 40, Issue 7
Consolidated bioprocessing of cellulosic biomass: an update
journal, October 2005
- Lynd, Lee R.; van Zyl, Willem H.; McBride, John E.
- Current Opinion in Biotechnology, Vol. 16, Issue 5, p. 577-583
A genome-scale Escherichia coli kinetic metabolic model k-ecoli457 satisfying flux data for multiple mutant strains
journal, December 2016
- Khodayari, Ali; Maranas, Costas D.
- Nature Communications, Vol. 7, Issue 1
Genome-scale metabolic analysis of Clostridium thermocellum for bioethanol production
journal, January 2010
- Roberts, Seth B.; Gowen, Christopher M.; Brooks, J. Paul
- BMC Systems Biology, Vol. 4, Issue 1, Article No. 31
Characterization of Nucleotide Pools as a Function of Physiological State in Escherichia coli
journal, October 2007
- Buckstein, M. H.; He, J.; Rubin, H.
- Journal of Bacteriology, Vol. 190, Issue 2
Consolidated Bioprocessing of Lignocellulosic Feedstocks for Ethanol Fuel Production
journal, December 2012
- Schuster, Brian G.; Chinn, Mari S.
- BioEnergy Research, Vol. 6, Issue 2
Transcarboxylase, ii. Purification and Properties of Methylmalonyl-Oxaloacetic Transcarboxylase
journal, March 1961
- Wood, H. G.; Stjernholm, R.
- Proceedings of the National Academy of Sciences, Vol. 47, Issue 3
Automated Genome Annotation and Metabolic Model Reconstruction in the SEED and Model SEED
book, January 2013
- Devoid, Scott; Overbeek, Ross; DeJongh, Matthew
- Methods in Molecular Biology
Redirecting carbon flux through exogenous pyruvate kinase to achieve high ethanol yields in Clostridium thermocellum
journal, January 2013
- Deng, Yu; Olson, Daniel G.; Zhou, Jilai
- Metabolic Engineering, Vol. 15, p. 151-158
Enzymological aspects of the pathways for trimethylamine oxidation and C1 assimilation in obligate methylotrophs and restricted facultative methylotrophs
journal, June 1975
- Colby, J.; Zatman, L. J.
- Biochemical Journal, Vol. 148, Issue 3
Engineering electron metabolism to increase ethanol production in Clostridium thermocellum
journal, January 2017
- Lo, Jonathan; Olson, Daniel G.; Murphy, Sean Jean-Loup
- Metabolic Engineering, Vol. 39
Changes in Transcript Abundance in Chlamydomonas reinhardtii following Nitrogen Deprivation Predict Diversion of Metabolism
journal, October 2010
- Miller, Rachel; Wu, Guangxi; Deshpande, Rahul R.
- Plant Physiology, Vol. 154, Issue 4
A hybrid of bees algorithm and flux balance analysis with OptKnock as a platform for in silico optimization of microbial strains
journal, July 2013
- Choon, Yee Wen; Mohamad, Mohd Saberi; Deris, Safaai
- Bioprocess and Biosystems Engineering, Vol. 37, Issue 3
OptForce: An Optimization Procedure for Identifying All Genetic Manipulations Leading to Targeted Overproductions
journal, April 2010
- Ranganathan, Sridhar; Suthers, Patrick F.; Maranas, Costas D.
- PLoS Computational Biology, Vol. 6, Issue 4
A kinetic model of Escherichia coli core metabolism satisfying multiple sets of mutant flux data
journal, September 2014
- Khodayari, Ali; Zomorrodi, Ali R.; Liao, James C.
- Metabolic Engineering, Vol. 25
Atypical Glycolysis in Clostridium thermocellum
journal, February 2013
- Zhou, Jilai; Olson, Daniel G.; Argyros, D. Aaron
- Applied and Environmental Microbiology, Vol. 79, Issue 9, p. 3000-3008
Improving prediction fidelity of cellular metabolism with kinetic descriptions
journal, December 2015
- Chowdhury, Anupam; Khodayari, Ali; Maranas, Costas D.
- Current Opinion in Biotechnology, Vol. 36
MetaboLights—an open-access general-purpose repository for metabolomics studies and associated meta-data
journal, October 2012
- Haug, Kenneth; Salek, Reza M.; Conesa, Pablo
- Nucleic Acids Research, Vol. 41, Issue D1
Likely features and costs of mature biomass ethanol technology
journal, March 1996
- Lynd, Lee R.; Elamder, Richard T.; Wyman, Charles E.
- Applied Biochemistry and Biotechnology, Vol. 57-58, Issue 1
Ensemble Modeling for Robustness Analysis in engineering non-native metabolic pathways
journal, September 2014
- Lee, Yun; Lafontaine Rivera, Jimmy G.; Liao, James C.
- Metabolic Engineering, Vol. 25
k-OptForce: Integrating Kinetics with Flux Balance Analysis for Strain Design
journal, February 2014
- Chowdhury, Anupam; Zomorrodi, Ali R.; Maranas, Costas D.
- PLoS Computational Biology, Vol. 10, Issue 2
BRENDA in 2015: exciting developments in its 25th year of existence
journal, November 2014
- Chang, Antje; Schomburg, Ida; Placzek, Sandra
- Nucleic Acids Research, Vol. 43, Issue D1
Purification, properties, and kinetic mechanism of coenzyme A-linked aldehyde dehydrogenase from Clostridium kluyveri
journal, September 1980
- Smith, Linda Tombras; Kaplan, Nathan O.
- Archives of Biochemistry and Biophysics, Vol. 203, Issue 2
Nutritional interdependence betweenThermoanaerobacter thermohydrosulfuricus andClostridium thermocellum
journal, August 1995
- Mori, Yutaka
- Archives of Microbiology, Vol. 164, Issue 2
Microbial Cellulose Utilization: Fundamentals and Biotechnology
journal, September 2002
- Lynd, L. R.; Weimer, P. J.; van Zyl, W. H.
- Microbiology and Molecular Biology Reviews, Vol. 66, Issue 3, p. 506-577
Applications of systems biology towards microbial fuel production
journal, October 2011
- Gowen, Christopher M.; Fong, Stephen S.
- Trends in Microbiology, Vol. 19, Issue 10
A kinetics modeling study on the inhibition of glucose on cellulosome of Clostridium thermocellum
journal, August 2015
- Zhang, Pengcheng; Wang, Buyun; Xiao, Qunfang
- Bioresource Technology, Vol. 190
Genome-scale metabolic modeling of a clostridial co-culture for consolidated bioprocessing
journal, July 2010
- Salimi, Fahimeh; Zhuang, Kai; Mahadevan, Radhakrishnan
- Biotechnology Journal, Vol. 5, Issue 7
Closing the carbon balance for fermentation by Clostridium thermocellum (ATCC 27405)
journal, January 2012
- Ellis, Lucas D.; Holwerda, Evert K.; Hogsett, David
- Bioresource Technology, Vol. 103, Issue 1
Mechanism of product inhibition for cellobiohydrolase Cel7A during hydrolysis of insoluble cellulose: Product Inhibition of Cellulase Cel7A
journal, January 2016
- Olsen, Johan P.; Alasepp, Kadri; Kari, Jeppe
- Biotechnology and Bioengineering, Vol. 113, Issue 6
Capturing the response of Clostridium acetobutylicumto chemical stressors using a regulated genome-scale metabolic model
journal, October 2014
- Dash, Satyakam; Mueller, Thomas J.; Venkataramanan, Keerthi P.
- Biotechnology for Biofuels, Vol. 7, Issue 1
Reassessment of the Transhydrogenase/Malate Shunt Pathway in Clostridium thermocellum ATCC 27405 through Kinetic Characterization of Malic Enzyme and Malate Dehydrogenase
journal, January 2015
- Taillefer, M.; Rydzak, T.; Levin, D. B.
- Applied and Environmental Microbiology, Vol. 81, Issue 7
Energy conservation via electron bifurcating ferredoxin reduction and proton/Na+ translocating ferredoxin oxidation
journal, February 2013
- Buckel, Wolfgang; Thauer, Rudolf K.
- Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1827, Issue 2
Elimination of hydrogenase active site assembly blocks H2 production and increases ethanol yield in Clostridium thermocellum
journal, January 2015
- Biswas, Ranjita; Zheng, Tianyong; Olson, Daniel G.
- Biotechnology for Biofuels, Vol. 8, Issue 1
Metabolic modeling of clostridia: current developments and applications
journal, January 2016
- Dash, Satyakam; Ng, Chiam Yu; Maranas, Costas D.
- FEMS Microbiology Letters, Vol. 363, Issue 4
The need for biofuels as part of a low carbon energy future
journal, June 2015
- Fulton, Lewis M.; Lynd, Lee R.; Körner, Alexander
- Biofuels, Bioproducts and Biorefining, Vol. 9, Issue 5
Stability of Ensemble Models Predicts Productivity of Enzymatic Systems
journal, March 2016
- Theisen, Matthew K.; Lafontaine Rivera, Jimmy G.; Liao, James C.
- PLOS Computational Biology, Vol. 12, Issue 3
Elimination of Thermodynamically Infeasible Loops in Steady-State Metabolic Models
journal, February 2011
- Schellenberger, Jan; Lewis, Nathan E.; Palsson, Bernhard Ø.
- Biophysical Journal, Vol. 100, Issue 3
Kinetic modeling of batch fermentation for Populus hydrolysate tolerant mutant and wild type strains of Clostridium thermocellum
journal, November 2013
- Linville, Jessica L.; Rodriguez, Miguel; Mielenz, Jonathan R.
- Bioresource Technology, Vol. 147
The exometabolome of Clostridium thermocellum reveals overflow metabolism at high cellulose loading
journal, October 2014
- Holwerda, Evert K.; Thorne, Philip G.; Olson, Daniel G.
- Biotechnology for Biofuels, Vol. 7, Issue 1
A defined growth medium with very low background carbon for culturing Clostridium thermocellum
journal, February 2012
- Holwerda, Evert K.; Hirst, Kyle D.; Lynd, Lee R.
- Journal of Industrial Microbiology & Biotechnology, Vol. 39, Issue 6
The cellulosome and cellulose degradation by anaerobic bacteria
journal, September 2001
- Schwarz, W. H.
- Applied Microbiology and Biotechnology, Vol. 56, Issue 5-6
What is flux balance analysis?
journal, March 2010
- Orth, Jeffrey D.; Thiele, Ines; Palsson, Bernhard Ø
- Nature Biotechnology, Vol. 28, Issue 3
Genome-scale model for Clostridium acetobutylicum : Part II. Development of specific proton flux states and numerically determined sub-systems
journal, December 2008
- Senger, Ryan S.; Papoutsakis, Eleftherios T.
- Biotechnology and Bioengineering, Vol. 101, Issue 5
Mutant selection and phenotypic and genetic characterization of ethanol-tolerant strains of Clostridium thermocellum
journal, August 2011
- Shao, Xiongjun; Raman, Babu; Zhu, Mingjun
- Applied Microbiology and Biotechnology, Vol. 92, Issue 3
Metabolic network reconstruction and genome-scale model of butanol-producing strain Clostridium beijerinckii NCIMB 8052
journal, January 2011
- Milne, Caroline B.; Eddy, James A.; Raju, Ravali
- BMC Systems Biology, Vol. 5, Issue 1
Works referencing / citing this record:
Assessing Escherichia coli metabolism models and simulation approaches in phenotype predictions: Validation against experimental data
journal, October 2018
- Costa, Rafael S.; Vinga, Susana
- Biotechnology Progress, Vol. 34, Issue 6
Pareto Optimality Explanation of the Glycolytic Alternatives in Nature
journal, February 2019
- Ng, Chiam Yu; Wang, Lin; Chowdhury, Anupam
- Scientific Reports, Vol. 9, Issue 1
CRISPR interference (CRISPRi) as transcriptional repression tool for Hungateiclostridium thermocellum DSM 1313
journal, August 2019
- Ganguly, Joyshree; Martin‐Pascual, Maria; Kranenburg, Richard
- Microbial Biotechnology, Vol. 13, Issue 2
From Escherichia coli mutant 13C labeling data to a core kinetic model: A kinetic model parameterization pipeline
journal, September 2019
- Foster, Charles J.; Gopalakrishnan, Saratram; Antoniewicz, Maciek R.
- PLOS Computational Biology, Vol. 15, Issue 9