Biotechnology for Biofuels
Thayer School of Engineering
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.
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.
Dash S, Khodayari A, Zhou J, Holwerda EK, Olson DG, Lynd LR, Maranas CD. Development of a core Clostridium thermocellum kinetic metabolic model consistent with multiple genetic perturbations. Biotechnol Biofuels. 2017 May 2;10:108. doi: 10.1186/s13068-017-0792-2. PMID: 28469704; PMCID: PMC5414155.
Dartmouth Digital Commons Citation
Dash, Satyakam; Khodayari, Ali; Zhou, Jilai; Holwerda, Evert K.; Olson, Daniel; Lynd, Lee; and Maranas, Costas, "Development of a Core Clostridium Thermocellum Kinetic Metabolic Model Consistent with Multiple Genetic Perturbations" (2017). Dartmouth Scholarship. 558.