Bio-fermentation of modified flavonoids: an example of in vivo diversification of secondary metabolites.
A bio-fermentation technique was used for the in vivo diversification of flavonoid structures based on expression in Escherichia coli of six O-methyltransferases (OMTs) from Mentha x piperita and one O-glucosyltransferase (GT) each from Arabidopsis thaliana and Allium cepa. Enzymes were shown to be regio-specific in in vitro experiments and modified a broad range of flavonoid substrates at various positions. Using the flavonol quercetin as a model substrate, we show that the product spectrum produced with the in vivo approach is identical to that found in vitro. Additionally, using mixed cultures of E. coli expressing different classes of modifying genes (OMTs and GTs), the production of polymethylated flavonoid glucosides was observed. This report demonstrates the potential to increase the structural diversity of plant secondary metabolites using a multi-enzyme, bio-fermentation approach.
Willits
Michael G
MG
Syngenta Biotechnology, Inc., 3054 Cornwallis Road, Research Triangle Park, NC 27709, USA.
Giovanni
Maïté
M
Prata
Rogerio T N
RT
Kramer
Catherine M
CM
De Luca
Vincenzo
V
Steffens
John C
JC
Graser
Gerson
G
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D005419:Flavonoids / Q000737:chemistry / Q000378:metabolism*; D005964:Glucosyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D036142:Mentha piperita / Q000201:enzymology; D008780:Methyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D019697:Onions / Q000201:enzymology; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013237:Stereoisomerism; D013379:Substrate Specificity
D005419:Flavonoids; D011994:Recombinant Proteins; D008780:Methyltransferases; D005964:Glucosyltransferases