Metabolic engineering of Saccharomyces cerevisiae for de novo production of dihydrochalcones with known antioxidant, antidiabetic, and sweet tasting properties.
Dihydrochalcones are plant secondary metabolites comprising molecules of significant commercial interest as antioxidants, antidiabetics, or sweeteners. To date, their heterologous biosynthesis in microorganisms has been achieved only by precursor feeding or as minor by-products in strains engineered for flavonoid production. Here, the native ScTSC13 was overexpressed in Saccharomyces cerevisiae to increase its side activity in reducing p-coumaroyl-CoA to p-dihydrocoumaroyl-CoA. De novo production of phloretin, the first committed dihydrochalcone, was achieved by co-expression of additional relevant pathway enzymes. Naringenin, a major by-product of the initial pathway, was practically eliminated by using a chalcone synthase from barley with unexpected substrate specificity. By further extension of the pathway from phloretin with decorating enzymes with known specificities for dihydrochalcones, and by exploiting substrate flexibility of enzymes involved in flavonoid biosynthesis, de novo production of the antioxidant molecule nothofagin, the antidiabetic molecule phlorizin, the sweet molecule naringin dihydrochalcone, and 3-hydroxyphloretin was achieved.
Evolva SA, Duggingerstrasse 23, 4153 Reinach, Switzerland; Department of Biology, Technical University Darmstadt, Schnittspahnstrasse 10, 64287 Darmstadt, Germany.
Michael
M
Eichenberger
Evolva Biotech A/S, Lersø Parkallé 42, 2100 Copenhagen, Denmark; Department of Science and Environment, Roskilde University, Universitetsvej 1, 4000 Roskilde, Denmark.
Beata Joanna
BJ
Lehka
Evolva SA, Duggingerstrasse 23, 4153 Reinach, Switzerland.
Christophe
C
Folly
Evolva SA, Duggingerstrasse 23, 4153 Reinach, Switzerland.
David
D
Fischer
Department of Food Quality and Nutrition, Fondazione Edmund Mach, Centro Ricerca e Innovazione, Via E. Mach 1, 38010 San Michele all'Adige (TN), Italy.
Stefan
S
Martens
Evolva SA, Duggingerstrasse 23, 4153 Reinach, Switzerland.
Ernesto
E
Simón
Evolva SA, Duggingerstrasse 23, 4153 Reinach, Switzerland. Electronic address: michaeln@evolva.com.
Michael
M
Naesby
D000975:Antioxidants / Q000008:administration & dosage; D053898:Biosynthetic Pathways / Q000502:physiology; D047188:Chalcones / Q000008:administration & dosage / Q000096:biosynthesis*; D024861:Genetic Enhancement / Q000379:methods; D007004:Hypoglycemic Agents / Q000008:administration & dosage / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D013549:Sweetening Agents / Q000008:administration & dosage / Q000378:metabolism*
D000975:Antioxidants; D047188:Chalcones; D007004:Hypoglycemic Agents; D029701:Saccharomyces cerevisiae Proteins; D013549:Sweetening Agents; C015812:dihydrochalcone
3-Hydroxyphloretin (PubChem CID: 11778945); Dihydrochalcone; Double bond reductase; Naringenin (PubChem CID: 932); Naringin dihydrochalcone; Naringin dihydrochalcone (PubChem CID: 9894584); Nothofagin; Nothofagin (PubChem CID: 42607691); Phloretin (PubChem CID: 4788); Phlorizin; Phlorizin (PubChem CID: 6072); Pinocembrin (PubChem CID: 68071); Saccharomyces cerevisiae; Trilobatin (PubChem CID: 6451798)