title: Identification of a bifunctional maize C- and O-glucosyltransferase.
abstract: Flavonoids accumulate in plant vacuoles usually as O-glycosylated derivatives, but several species can also synthesize flavonoid C-glycosides. Recently, we demonstrated that a flavanone 2-hydroxylase (ZmF2H1, CYP93G5) converts flavanones to the corresponding 2-hydroxy derivatives, which are expected to serve as substrates for C-glycosylation. Here, we isolated a cDNA encoding a UDP-dependent glycosyltransferase (UGT708A6), and its activity was characterized by in vitro and in vivo bioconversion assays. In vitro assays using 2-hydroxyflavanones as substrates and in vivo activity assays in yeast co-expressing ZmF2H1 and UGT708A6 show the formation of the flavones C-glycosides. UGT708A6 can also O-glycosylate flavanones in bioconversion assays in Escherichia coli as well as by in vitro assays with the purified recombinant protein. Thus, UGT708A6 is a bifunctional glycosyltransferase that can produce both C- and O-glycosidated flavonoids, a property not previously described for any other glycosyltransferase.
authors.affiliation: From the Centro de Estudios Fotosintéticos y Bioquímicos.
authors.forename: María Lorena
authors.initials: ML
authors.lastname: Falcone Ferreyra
authors.forename: Eduardo
authors.initials: E
authors.lastname: Rodriguez
authors.forename: María Isabel
authors.initials: MI
authors.lastname: Casas
authors.forename: Guillermo
authors.initials: G
authors.lastname: Labadie
authors.forename: Erich
authors.initials: E
authors.lastname: Grotewold
authors.forename: Paula
authors.initials: P
authors.lastname: Casati
mesh_terms: D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D047309:Flavones / Q000235:genetics / Q000378:metabolism; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism*; D006027:Glycosides / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D003313:Zea mays / Q000201:enzymology* / Q000235:genetics
chemical_list: D047309:Flavones; D006027:Glycosides; D010940:Plant Proteins; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System; D005964:Glucosyltransferases
keywords: Flavonoids; Glycosylation; Glycosyltransferases; Maysin; Metabolic Engineering; Plant Biochemistry
references.citation: J Nat Prod. 2003 Sep;66(9):1280-3
references.pmid: 14510618
references.citation: Plant J. 1994 Jan;5(1):69-80
references.pmid: 8130799
references.citation: J Agric Food Chem. 2009 Sep 23;57(18):8142-60
references.pmid: 19719126
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: J Biol Chem. 2009 Jul 3;284(27):17926-34
references.pmid: 19411659
references.citation: Methods Enzymol. 2002;350:87-96
references.pmid: 12073338
references.citation: Proc Natl Acad Sci U S A. 1998 Mar 3;95(5):1996-2000
references.pmid: 9482823
references.citation: Mol Biol Evol. 2007 Aug;24(8):1596-9
references.pmid: 17488738
references.citation: Plant Cell. 2012 Jul;24(7):2745-64
references.pmid: 22822204
references.citation: Bioorg Med Chem. 2008 Sep 1;16(17):8117-26
references.pmid: 18682327
references.citation: Genetics. 1988 May;119(1):185-97
references.pmid: 3396861
references.citation: Plant Mol Biol. 1988 Jul;11(4):473-81
references.pmid: 24272404
references.citation: Front Plant Sci. 2012 Sep 28;3:222
references.pmid: 23060891
references.citation: Plant Cell. 1998 May;10(5):721-40
references.pmid: 9596632
references.citation: Methods Mol Biol. 2000;132:365-86
references.pmid: 10547847
references.citation: Annu Rev Plant Biol. 2006;57:761-80
references.pmid: 16669781
references.citation: Sci Pharm. 2011;79(4):963-75
references.pmid: 22145118
references.citation: Plant J. 2011 May;66(4):553-63
references.pmid: 21299659
references.citation: Curr Opin Plant Biol. 2005 Jun;8(3):254-63
references.pmid: 15860422
references.citation: Gene. 1988 Dec 30;74(2):527-34
references.pmid: 3073106
references.citation: Plant Physiol. 2007 Jun;144(2):741-51
references.pmid: 17434990
references.citation: J Exp Bot. 2012 May;63(9):3429-44
references.pmid: 22213816
references.citation: Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):8820-5
references.pmid: 11607699
references.citation: Proc Natl Acad Sci U S A. 2007 Dec 18;104(51):20238-43
references.pmid: 18077347
references.citation: Molecules. 2010 Nov 16;15(11):8251-9
references.pmid: 21081855
references.citation: Nat Chem Biol. 2007 Jul;3(7):352
references.pmid: 17576414
references.citation: BMC Plant Biol. 2009 Dec 29;9:151
references.pmid: 20040108
references.citation: Plant J. 2012 Mar;69(6):1030-42
references.pmid: 22077743
references.citation: Plant Cell. 2013 May;25(5):1840-54
references.pmid: 23723325
references.citation: Biotechniques. 2003 Feb;34(2):374-8
references.pmid: 12613259
references.citation: Photochem Photobiol. 2007 Sep-Oct;83(5):1167-73
references.pmid: 17880511
references.citation: J Biol Chem. 1997 Aug 1;272(31):19176-86
references.pmid: 9235908
references.citation: J Hered. 2004 May-Jun;95(3):225-33
references.pmid: 15220389
references.citation: Phytochemistry. 2009 Feb;70(3):325-47
references.pmid: 19217634
references.citation: Rapid Commun Mass Spectrom. 2008 Jun;22(12):1834-42
references.pmid: 18470889
references.citation: Angew Chem Int Ed Engl. 2004 May 24;43(22):2962-5
references.pmid: 15170316
references.citation: FEBS Lett. 1998 Jul 17;431(2):287-90
references.pmid: 9708921
references.citation: Annu Rev Biochem. 2008;77:521-55
references.pmid: 18518825
references.citation: Appl Environ Microbiol. 2005 May;71(5):2539-47
references.pmid: 15870344
references.citation: Plant Physiol. 2010 Sep;154(1):324-33
references.pmid: 20647377
references.citation: Metab Eng. 2013 Mar;16:11-20
references.pmid: 23246521
references.citation: Plant Cell. 2005 Nov;17(11):3141-54
references.pmid: 16214900
references.citation: Plant Physiol. 2010 Nov;154(3):1024-39
references.pmid: 20833728
references.citation: Crit Rev Food Sci Nutr. 2006;46(2):161-83
references.pmid: 16431408