title: Genome-scale modeling enables metabolic engineering of Saccharomyces cerevisiae for succinic acid production.
abstract: In this work, we describe the application of a genome-scale metabolic model and flux balance analysis for the prediction of succinic acid overproduction strategies in Saccharomyces cerevisiae. The top three single gene deletion strategies, Δmdh1, Δoac1, and Δdic1, were tested using knock-out strains cultivated anaerobically on glucose, coupled with physiological and DNA microarray characterization. While Δmdh1 and Δoac1 strains failed to produce succinate, Δdic1 produced 0.02 C-mol/C-mol glucose, in close agreement with model predictions (0.03 C-mol/C-mol glucose). Transcriptional profiling suggests that succinate formation is coupled to mitochondrial redox balancing, and more specifically, reductive TCA cycle activity. While far from industrial titers, this proof-of-concept suggests that in silico predictions coupled with experimental validation can be used to identify novel and non-intuitive metabolic engineering strategies.
authors.affiliation: Department of Chemical and Biological Engineering, Chalmers University of Technology, SE-41296, Gothenburg, Sweden.
authors.forename: Rasmus
authors.initials: R
authors.lastname: Agren
authors.forename: José Manuel
authors.initials: JM
authors.lastname: Otero
authors.forename: Jens
authors.initials: J
authors.lastname: Nielsen
mesh_terms: D000693:Anaerobiosis; D002952:Citric Acid Cycle / Q000235:genetics; D003198:Computer Simulation; D005285:Fermentation; D017353:Gene Deletion; D005800:Genes, Fungal / Q000235:genetics; D016681:Genome, Fungal / Q000235:genetics*; D005947:Glucose / Q000378:metabolism; D060847:Metabolic Engineering; D008928:Mitochondria / Q000378:metabolism; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis; D010084:Oxidation-Reduction; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D019802:Succinic Acid / Q000378:metabolism*; D059467:Transcriptome
chemical_list: D019802:Succinic Acid; D005947:Glucose