skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: A simple method for enzymatic synthesis of unlabeled and radiolabeled Hydroxycinnamate-CoA

Abstract

Hydroxycinnamate coenzyme A (CoA) thioesters are substrates for biosynthesis of lignin and hydroxycinna- mate esters of polysaccharides and other polymers. Hence, a supply of these substrates is essential for investigation of cell wall biosynthesis. In this study, three recombinant enzymes, caffeic acid 3-O-methyltransferase, 4-coumarate- CoA ligase 1, and 4-coumarate-CoA ligase 5, were cloned from wheat, tobacco, and Arabidopsis, respectively, and were used to synthesize {sup 14}C-feruloyl-CoA, caffeoyl-CoA, p-coumaroyl-CoA, feruloyl-CoA, and sinapoyl-CoA. The corresponding hydroxycinnamoyl-CoA thioesters were high-performance liquid chromatography purified, the only extraction/purification step necessary, with total yields between 88-95%. Radiolabeled {sup 14}C-feruloyl-CoA was generated from caffeic acid and S-adenosyl-{sup 14}C-methionine under the combined action of caffeic acid 3-O-methyltransferase and 4-coumarate-CoA ligase 1. About 70% of {sup 14}C-methyl groups from S-adenosyl methionine were incorporated into the final product. The methods presented are simple, fast, and efficient for the preparation of the hydroxycinnamate thioesters.

Authors:
; ; ;
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
Physical Biosciences Division
OSTI Identifier:
1051785
Report Number(s):
LBNL-5185E
Journal ID: ISSN 1939-1234
DOE Contract Number:  
DE-AC02-05CH11231
Resource Type:
Journal Article
Journal Name:
BioEnergy Research
Additional Journal Information:
Journal Volume: 3; Journal Issue: 2; Related Information: Journal Publication Date: 30 MAR 2010; Journal ID: ISSN 1939-1234
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; 4-Coumarate-CoA ligase, Caffeoyl O-methyltransferase, Cell walls, Hydroxycinnamates, Ferulic acid

Citation Formats

Rautergarten, Carsten, Baidoo, Edward, Keasling, Jay, and Vibe Scheller, Henrik. A simple method for enzymatic synthesis of unlabeled and radiolabeled Hydroxycinnamate-CoA. United States: N. p., 2011. Web. doi:10.1007/s12155-010-9085-3.
Rautergarten, Carsten, Baidoo, Edward, Keasling, Jay, & Vibe Scheller, Henrik. A simple method for enzymatic synthesis of unlabeled and radiolabeled Hydroxycinnamate-CoA. United States. doi:10.1007/s12155-010-9085-3.
Rautergarten, Carsten, Baidoo, Edward, Keasling, Jay, and Vibe Scheller, Henrik. Wed . "A simple method for enzymatic synthesis of unlabeled and radiolabeled Hydroxycinnamate-CoA". United States. doi:10.1007/s12155-010-9085-3. https://www.osti.gov/servlets/purl/1051785.
@article{osti_1051785,
title = {A simple method for enzymatic synthesis of unlabeled and radiolabeled Hydroxycinnamate-CoA},
author = {Rautergarten, Carsten and Baidoo, Edward and Keasling, Jay and Vibe Scheller, Henrik},
abstractNote = {Hydroxycinnamate coenzyme A (CoA) thioesters are substrates for biosynthesis of lignin and hydroxycinna- mate esters of polysaccharides and other polymers. Hence, a supply of these substrates is essential for investigation of cell wall biosynthesis. In this study, three recombinant enzymes, caffeic acid 3-O-methyltransferase, 4-coumarate- CoA ligase 1, and 4-coumarate-CoA ligase 5, were cloned from wheat, tobacco, and Arabidopsis, respectively, and were used to synthesize {sup 14}C-feruloyl-CoA, caffeoyl-CoA, p-coumaroyl-CoA, feruloyl-CoA, and sinapoyl-CoA. The corresponding hydroxycinnamoyl-CoA thioesters were high-performance liquid chromatography purified, the only extraction/purification step necessary, with total yields between 88-95%. Radiolabeled {sup 14}C-feruloyl-CoA was generated from caffeic acid and S-adenosyl-{sup 14}C-methionine under the combined action of caffeic acid 3-O-methyltransferase and 4-coumarate-CoA ligase 1. About 70% of {sup 14}C-methyl groups from S-adenosyl methionine were incorporated into the final product. The methods presented are simple, fast, and efficient for the preparation of the hydroxycinnamate thioesters.},
doi = {10.1007/s12155-010-9085-3},
journal = {BioEnergy Research},
issn = {1939-1234},
number = 2,
volume = 3,
place = {United States},
year = {2011},
month = {7}
}