Expression of a soluble flavone synthase allows the biosynthesis of phytoestrogen derivatives in Escherichia coli.
Flavones are plant secondary metabolites with potent pharmacological properties. We report the functional expression of FSI, a flavonoid 2-oxoglutarate-dependent dioxygenase-encoding flavone synthase from parsley in Escherichia coli. This expression allows the biosynthesis of various flavones from phenylpropanoid acids in recombinant E. coli strains simultaneously expressing five plant-specific flavone biosynthetic genes. The gene ensemble consists of 4CL-2 (4-coumarate:CoA ligase) and FSI (flavone synthase I) from parsley, chsA (chalcone synthase) and chiA (chalcone isomerase) from Petunia hybrida, and OMT1A (7-O-methyltransferase) from peppermint. After a 24-h cultivation, the recombinant E. coli produces significant amounts of apigenin (415 microg/l), luteolin (10 microg/l), and genkwanin (208 microg/l). The majority of the flavone products are excreted in the culture media; however, 25% is contained within the cells. The metabolic engineering strategy presented demonstrates that plant-specific flavones are successfully produced in E. coli for the first time by incorporating a soluble flavone synthase confined only in Apiaceae.
Leonard
Effendi
E
Department of Chemical and Biological Engineering, University at Buffalo, the State University of New York, 904 Furnas Hall, Buffalo, NY 14260, USA. mkoffas@buffalo.edu
Chemler
Joseph
J
Lim
Kok Hong
KH
Koffas
Mattheos A G
MA
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D047309:Flavones / Q000737:chemistry / Q000378:metabolism; D015870:Gene Expression; D005818:Genetic Engineering; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism*; D015394:Molecular Structure; D028528:Petroselinum / Q000201:enzymology / Q000235:genetics; D048789:Phytoestrogens / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D012995:Solubility
D047309:Flavones; D048789:Phytoestrogens; D010940:Plant Proteins; D006899:Mixed Function Oxygenases; C416048:flavonol 6-hydroxylase