By the same authors

From the same journal

From the same journal

Regioselectivity of glucosylation of caffeic acid by a UDP-glucose : glucosyltransferase is maintained in planta

Research output: Contribution to journalArticle

Standard

Regioselectivity of glucosylation of caffeic acid by a UDP-glucose : glucosyltransferase is maintained in planta. / Lim, E K; Higgins, G S; Li, Y; Bowles, D J.

In: Biochemical journal, Vol. 373, 01.08.2003, p. 987-992.

Research output: Contribution to journalArticle

Harvard

Lim, EK, Higgins, GS, Li, Y & Bowles, DJ 2003, 'Regioselectivity of glucosylation of caffeic acid by a UDP-glucose : glucosyltransferase is maintained in planta', Biochemical journal, vol. 373, pp. 987-992. https://doi.org/10.1042/BJ20021453

APA

Lim, E. K., Higgins, G. S., Li, Y., & Bowles, D. J. (2003). Regioselectivity of glucosylation of caffeic acid by a UDP-glucose : glucosyltransferase is maintained in planta. Biochemical journal, 373, 987-992. https://doi.org/10.1042/BJ20021453

Vancouver

Lim EK, Higgins GS, Li Y, Bowles DJ. Regioselectivity of glucosylation of caffeic acid by a UDP-glucose : glucosyltransferase is maintained in planta. Biochemical journal. 2003 Aug 1;373:987-992. https://doi.org/10.1042/BJ20021453

Author

Lim, E K ; Higgins, G S ; Li, Y ; Bowles, D J. / Regioselectivity of glucosylation of caffeic acid by a UDP-glucose : glucosyltransferase is maintained in planta. In: Biochemical journal. 2003 ; Vol. 373. pp. 987-992.

Bibtex - Download

@article{f73418e1df2a4b098d4cdf5d2dfef6b0,
title = "Regioselectivity of glucosylation of caffeic acid by a UDP-glucose : glucosyltransferase is maintained in planta",
abstract = "Caffeic acid is a phenylpropanoid playing an important role in the pathways leading to lignin synthesis and the production of a wide variety of secondary metabolites. The compound is also an antioxidant and has potential utility as a general protectant against free radicals. Three glucosylated forms of caffeic acid are known to exist: the 3-O- and 4-O-glucosides and the glucose ester. This study describes for the first time a glucosyltransferase [UDP-glucose:glucosyltransferase (UGT)] that is specific for the 3-hydroxyl, and not the 4-hydroxyl, position of caffeic acid. The UGT sequence of Arabidopsis, UGT71C1, has been expressed as a recombinant fusion protein in Escherichia coli, purified and assayed against a range of substrates in vitro. The assay confirmed that caffeic acid as the preferred substrate when compared with other hydroxycinnamates, although UGT71C1 also exhibited substantial activity towards flavonoid substrates, known to have structural features that can be recognized by many different UGTs. The expression of UGT71C1 in transgenic Arabidopsis was driven by the constitutive cauliflower mosaic virus 35 S (CaMV35S) promoter. Nine independent transgenic lines were taken to homozygosity and characterized by Northern-blot analysis, assay of enzyme activity in leaf extracts and HPLC analysis of the glucosides. The level of expression of UGT71C1 was enhanced considerably in several lines, leading to a higher level of the corresponding enzyme activity and a higher level of caffeoyl-3-0-glucoside. The data are discussed in the context of the utility of UGTs for natural product biotransformations.",
keywords = "Arabidopsis family 1 glycosyltransferase, overexpression, recombinant protein, target profiling, RADICAL-SCAVENGING ACTIVITY, BIOCHEMICAL-CHARACTERIZATION, ARABIDOPSIS-THALIANA, SUSPENSION-CULTURES, LIGNIN, ESTER, IDENTIFICATION, BIOSYNTHESIS, OXIDATION, PEROXYL",
author = "Lim, {E K} and Higgins, {G S} and Y Li and Bowles, {D J}",
year = "2003",
month = "8",
day = "1",
doi = "10.1042/BJ20021453",
language = "English",
volume = "373",
pages = "987--992",
journal = "Biochemical journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Regioselectivity of glucosylation of caffeic acid by a UDP-glucose : glucosyltransferase is maintained in planta

AU - Lim, E K

AU - Higgins, G S

AU - Li, Y

AU - Bowles, D J

PY - 2003/8/1

Y1 - 2003/8/1

N2 - Caffeic acid is a phenylpropanoid playing an important role in the pathways leading to lignin synthesis and the production of a wide variety of secondary metabolites. The compound is also an antioxidant and has potential utility as a general protectant against free radicals. Three glucosylated forms of caffeic acid are known to exist: the 3-O- and 4-O-glucosides and the glucose ester. This study describes for the first time a glucosyltransferase [UDP-glucose:glucosyltransferase (UGT)] that is specific for the 3-hydroxyl, and not the 4-hydroxyl, position of caffeic acid. The UGT sequence of Arabidopsis, UGT71C1, has been expressed as a recombinant fusion protein in Escherichia coli, purified and assayed against a range of substrates in vitro. The assay confirmed that caffeic acid as the preferred substrate when compared with other hydroxycinnamates, although UGT71C1 also exhibited substantial activity towards flavonoid substrates, known to have structural features that can be recognized by many different UGTs. The expression of UGT71C1 in transgenic Arabidopsis was driven by the constitutive cauliflower mosaic virus 35 S (CaMV35S) promoter. Nine independent transgenic lines were taken to homozygosity and characterized by Northern-blot analysis, assay of enzyme activity in leaf extracts and HPLC analysis of the glucosides. The level of expression of UGT71C1 was enhanced considerably in several lines, leading to a higher level of the corresponding enzyme activity and a higher level of caffeoyl-3-0-glucoside. The data are discussed in the context of the utility of UGTs for natural product biotransformations.

AB - Caffeic acid is a phenylpropanoid playing an important role in the pathways leading to lignin synthesis and the production of a wide variety of secondary metabolites. The compound is also an antioxidant and has potential utility as a general protectant against free radicals. Three glucosylated forms of caffeic acid are known to exist: the 3-O- and 4-O-glucosides and the glucose ester. This study describes for the first time a glucosyltransferase [UDP-glucose:glucosyltransferase (UGT)] that is specific for the 3-hydroxyl, and not the 4-hydroxyl, position of caffeic acid. The UGT sequence of Arabidopsis, UGT71C1, has been expressed as a recombinant fusion protein in Escherichia coli, purified and assayed against a range of substrates in vitro. The assay confirmed that caffeic acid as the preferred substrate when compared with other hydroxycinnamates, although UGT71C1 also exhibited substantial activity towards flavonoid substrates, known to have structural features that can be recognized by many different UGTs. The expression of UGT71C1 in transgenic Arabidopsis was driven by the constitutive cauliflower mosaic virus 35 S (CaMV35S) promoter. Nine independent transgenic lines were taken to homozygosity and characterized by Northern-blot analysis, assay of enzyme activity in leaf extracts and HPLC analysis of the glucosides. The level of expression of UGT71C1 was enhanced considerably in several lines, leading to a higher level of the corresponding enzyme activity and a higher level of caffeoyl-3-0-glucoside. The data are discussed in the context of the utility of UGTs for natural product biotransformations.

KW - Arabidopsis family 1 glycosyltransferase

KW - overexpression

KW - recombinant protein

KW - target profiling

KW - RADICAL-SCAVENGING ACTIVITY

KW - BIOCHEMICAL-CHARACTERIZATION

KW - ARABIDOPSIS-THALIANA

KW - SUSPENSION-CULTURES

KW - LIGNIN

KW - ESTER

KW - IDENTIFICATION

KW - BIOSYNTHESIS

KW - OXIDATION

KW - PEROXYL

U2 - 10.1042/BJ20021453

DO - 10.1042/BJ20021453

M3 - Article

VL - 373

SP - 987

EP - 992

JO - Biochemical journal

JF - Biochemical journal

SN - 0264-6021

ER -