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

Journal Article · Wed Jul 20 04:00:00 EDT 2011 · BioEnergy Research

DOI:https://doi.org/10.1007/s12155-010-9085-3· OSTI ID:1051785

Rautergarten, Carsten; Baidoo, Edward; Keasling, Jay; Vibe Scheller, Henrik

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.

• Research Organization: Ernest Orlando Lawrence Berkeley National Laboratory, Berkeley, CA (US)
• Sponsoring Organization: Physical Biosciences Division
• DOE Contract Number: AC02-05CH11231
• OSTI ID: 1051785
• Report Number(s): LBNL-5185E
• Journal Information: BioEnergy Research, Journal Name: BioEnergy Research Journal Issue: 2 Vol. 3; ISSN 1939-1234
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
4-Coumarate-CoA ligase
Caffeoyl O-methyltransferase
Cell walls
Hydroxycinnamates
Ferulic acid
59 BASIC BIOLOGICAL SCIENCES