title: Model-aided atpE gene knockout strategy in Escherichia coli for enhanced succinic acid production from glycerol.
abstract: Succinic acid is an important platform chemical with a variety of applications. Model-guided metabolic engineering strategies in Escherichia coli for strain improvement to increase succinic acid production using glucose and glycerol remain largely unexplored. Herein, we report what are, to our knowledge, the first metabolic knockout of the atpE gene to have increased succinic acid production using both glucose and alternative glycerol carbon sources in E. coli. Guided by a genome-scale metabolic model, we engineered the E. coli host to enhance anaerobic production of succinic acid by deleting the atpE gene, thereby generating additional reducing equivalents by blocking H(+) conduction across the mutant cell membrane. This strategy produced 1.58 and .49 g l(-1) of succinic acid from glycerol and glucose substrate, respectively. This work further elucidates a model-guided and/or system-based metabolic engineering, involving only a single-gene deletion strategy for enhanced succinic acid production in E. coli.
authors.affiliation: a Bioinformatics Research Group (BIRG), Faculty of Biosciences and Medical Engineering, Department of Biosciences and Health Sciences , Universiti Teknologi Malaysia , 81310 Skudai , Johor Bahru , Malaysia.
authors.forename: Bashir Sajo
authors.initials: BS
authors.lastname: Mienda
authors.affiliation: a Bioinformatics Research Group (BIRG), Faculty of Biosciences and Medical Engineering, Department of Biosciences and Health Sciences , Universiti Teknologi Malaysia , 81310 Skudai , Johor Bahru , Malaysia.
authors.forename: Mohd Shahir
authors.initials: MS
authors.lastname: Shamsir
authors.affiliation: b Faculty of Chemical Engineering, Department of Bioprocess Engineering , Universiti Teknologi Malaysia , 81310 Skudai , Johor Bahru , Malaysia.
authors.forename: Rosli
authors.initials: R
authors.lastname: Md Illias
mesh_terms: D025243:Bacterial Proton-Translocating ATPases / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism*; D005285:Fermentation; D055786:Gene Knockout Techniques; D005947:Glucose / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D053858:Metabolic Networks and Pathways; D008954:Models, Biological; D009154:Mutation; D019802:Succinic Acid / Q000378:metabolism*
chemical_list: D029968:Escherichia coli Proteins; C074429:atpE protein, E coli; D019802:Succinic Acid; D025243:Bacterial Proton-Translocating ATPases; D005947:Glucose; D005990:Glycerol
keywords: E. coli; F0 sector of membrane-bound ATP synthase (atpE); OptFlux; gene knockout; glycerol; metabolic engineering; model-guided; succinic acid