Optimal cofactor swapping can increase the theoretical yield for chemical production in Escherichia coli and Saccharomyces cerevisiae.
Maintaining cofactor balance is a critical function in microorganisms, but often the native cofactor balance does not match the needs of an engineered metabolic flux state. Here, an optimization procedure is utilized to identify optimal cofactor-specificity "swaps" for oxidoreductase enzymes utilizing NAD(H) or NADP(H) in the genome-scale metabolic models of Escherichia coli and Saccharomyces cerevisiae. The theoretical yields of all native carbon-containing molecules are considered, as well as theoretical yields of twelve heterologous production pathways in E. coli. Swapping the cofactor specificity of central metabolic enzymes (especially GAPD and ALCD2x) is shown to increase NADPH production and increase theoretical yields for native products in E. coli and yeast--including L-aspartate, L-lysine, L-isoleucine, L-proline, L-serine, and putrescine--and non-native products in E. coli-including 1,3-propanediol, 3-hydroxybutyrate, 3-hydroxypropanoate, 3-hydroxyvalerate, and styrene.
Department of Bioengineering, University of California, 9500 Gilman Drive #0412, San Diego, La Jolla, CA 92093-0412, USA.
Zachary A
ZA
King
Department of Bioengineering, University of California, 9500 Gilman Drive #0412, San Diego, La Jolla, CA 92093-0412, USA; Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Lyngby, Denmark. Electronic address: afeist@ucsd.edu.
Adam M
AM
Feist
D020155:3-Hydroxybutyric Acid / Q000096:biosynthesis; D000596:Amino Acids / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D019344:Lactic Acid / Q000031:analogs & derivatives / Q000096:biosynthesis; D009243:NAD / Q000235:genetics / Q000378:metabolism; D009249:NADP / Q000235:genetics / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D010421:Pentanoic Acids / Q000378:metabolism; D011409:Propylene Glycols / Q000378:metabolism; D011700:Putrescine / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D020058:Styrene / Q000378:metabolism
D000596:Amino Acids; D029968:Escherichia coli Proteins; D010421:Pentanoic Acids; D011409:Propylene Glycols; D029701:Saccharomyces cerevisiae Proteins; D009243:NAD; C013056:beta-hydroxyvaleric acid; D019344:Lactic Acid; D020058:Styrene; D009249:NADP; C041787:1,3-propanediol; C031601:hydracrylic acid; D010088:Oxidoreductases; D020155:3-Hydroxybutyric Acid; D011700:Putrescine
Cofactor balancing; Constraint-based modeling; Escherichia coli; MILP; Metabolic engineering; Saccharomyces cerevisiae; Theoretical yield