Identification of UGT84A13 as a candidate enzyme for the first committed step of gallotannin biosynthesis in pedunculate oak (Quercus robur)

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Identification of UGT84A13 as a candidate enzyme for the first committed step of gallotannin biosynthesis in pedunculate oak (Quercus robur). / Mittasch, Juliane; Böttcher, Christoph; Frolova, Nadezhda; Bönn, Markus; Milkowski, Carsten.

в: Phytochemistry, Том 99, 2014, стр. 44-51.

Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование

BibTeX

@article{9513ae8477154757b48bfc4deb486c24,

title = "Identification of UGT84A13 as a candidate enzyme for the first committed step of gallotannin biosynthesis in pedunculate oak (Quercus robur)",

abstract = "A cDNA encoding the ester-forming hydroxybenzoic acid glucosyltransferase UGT84A13 was isolated from a cDNA library of Quercus robur swelling buds and young leaves. The enzyme displayed high sequence identity to resveratrol/hydroxycinnamate and hydroxybenzoate/hydroxycinnamate glucosyltransferases from Vitis species and clustered to the phylogenetic group L of plant glucosyltransferases, mainly involved in the formation of 1-O-β-D-glucose esters. In silico transcriptome analysis confirmed expression of UGT84A13 in Quercus tissues which were previously shown to exhibit UDP-glucose:gallic acid glucosyltransferase activity. UGT84A13 was functionally expressed in Escherichia coli as N-terminal His-tagged protein. In vitro kinetic measurements with the purified recombinant enzyme revealed a clear preference for hydroxybenzoic acids as glucosyl acceptor in comparison to hydroxycinnamic acids. Of the preferred in vitro substrates, protocatechuic, vanillic and gallic acid, only the latter and its corresponding 1-O-{\ss}-D-glucose ester were found to be accumulated in young oak leaves. This indicates that in planta UGT84A13 catalyzes the formation of , 1-O-galloyl-{\ss}-D-glucose, the first committed step of gallotannin biosynthesis.",

keywords = "Glucosyltransferases/isolation & purification, Hydrolyzable Tannins/chemistry, Molecular Structure, Plant Leaves/enzymology, Plant Shoots/enzymology, Quercus/enzymology",

author = "Juliane Mittasch and Christoph B{\"o}ttcher and Nadezhda Frolova and Markus B{\"o}nn and Carsten Milkowski",

year = "2014",

doi = "10.1016/j.phytochem.2013.11.023",

language = "English",

volume = "99",

pages = "44--51",

journal = "Phytochemistry",

issn = "0031-9422",

publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Identification of UGT84A13 as a candidate enzyme for the first committed step of gallotannin biosynthesis in pedunculate oak (Quercus robur)

AU - Mittasch, Juliane

AU - Böttcher, Christoph

AU - Frolova, Nadezhda

AU - Bönn, Markus

AU - Milkowski, Carsten

PY - 2014

Y1 - 2014

N2 - A cDNA encoding the ester-forming hydroxybenzoic acid glucosyltransferase UGT84A13 was isolated from a cDNA library of Quercus robur swelling buds and young leaves. The enzyme displayed high sequence identity to resveratrol/hydroxycinnamate and hydroxybenzoate/hydroxycinnamate glucosyltransferases from Vitis species and clustered to the phylogenetic group L of plant glucosyltransferases, mainly involved in the formation of 1-O-β-D-glucose esters. In silico transcriptome analysis confirmed expression of UGT84A13 in Quercus tissues which were previously shown to exhibit UDP-glucose:gallic acid glucosyltransferase activity. UGT84A13 was functionally expressed in Escherichia coli as N-terminal His-tagged protein. In vitro kinetic measurements with the purified recombinant enzyme revealed a clear preference for hydroxybenzoic acids as glucosyl acceptor in comparison to hydroxycinnamic acids. Of the preferred in vitro substrates, protocatechuic, vanillic and gallic acid, only the latter and its corresponding 1-O-ß-D-glucose ester were found to be accumulated in young oak leaves. This indicates that in planta UGT84A13 catalyzes the formation of , 1-O-galloyl-ß-D-glucose, the first committed step of gallotannin biosynthesis.

AB - A cDNA encoding the ester-forming hydroxybenzoic acid glucosyltransferase UGT84A13 was isolated from a cDNA library of Quercus robur swelling buds and young leaves. The enzyme displayed high sequence identity to resveratrol/hydroxycinnamate and hydroxybenzoate/hydroxycinnamate glucosyltransferases from Vitis species and clustered to the phylogenetic group L of plant glucosyltransferases, mainly involved in the formation of 1-O-β-D-glucose esters. In silico transcriptome analysis confirmed expression of UGT84A13 in Quercus tissues which were previously shown to exhibit UDP-glucose:gallic acid glucosyltransferase activity. UGT84A13 was functionally expressed in Escherichia coli as N-terminal His-tagged protein. In vitro kinetic measurements with the purified recombinant enzyme revealed a clear preference for hydroxybenzoic acids as glucosyl acceptor in comparison to hydroxycinnamic acids. Of the preferred in vitro substrates, protocatechuic, vanillic and gallic acid, only the latter and its corresponding 1-O-ß-D-glucose ester were found to be accumulated in young oak leaves. This indicates that in planta UGT84A13 catalyzes the formation of , 1-O-galloyl-ß-D-glucose, the first committed step of gallotannin biosynthesis.

KW - Glucosyltransferases/isolation & purification

KW - Hydrolyzable Tannins/chemistry

KW - Molecular Structure

KW - Plant Leaves/enzymology

KW - Plant Shoots/enzymology

KW - Quercus/enzymology

U2 - 10.1016/j.phytochem.2013.11.023

DO - 10.1016/j.phytochem.2013.11.023

M3 - Article

C2 - 24412325

VL - 99

SP - 44

EP - 51

JO - Phytochemistry

JF - Phytochemistry

SN - 0031-9422

ER -

ID: 5715767