Abstract
Hydroxycinnamoyltransferases (HCTs) catalyze the transfer of the cinnamoyl moiety from hydroxycinnamoyl-CoA to various acceptors such as shikimic acid, quinic acid, hydroxylated acid, and glycerol. Four rice HCT homologues (OsHCT1-4) to tobacco HST were cloned, and OsHCT4 was expressed in Escherichia coli as a glutathione S-transferase fusion protein. Using the purified recombinant protein and biotransformation techniques, whether OsHCT4 shows hydroxycinnamoyltransferase activity with a variety of acyl group acceptors was investigated. The results of high performance liquid chromatography (HPLC) and mass spectrometry (MS) established that OsHCT4 mediated the trans-esterification of glycerol as well as shikimic acid in the presence of hydroxycinnamoyl-CoA. The structure of the reaction product was determined using nuclear magnetic resonance spectroscopy (NMR). E. coli cells co-expressing 4CL (4-coumarate:coenzyme A ligase) and OsHCT4 converted p-coumaric acid, ferulic acid, and caffeic acid into the corresponding glycerides. While this conversion is very efficient in vitro, the physiological significant in rice is currently unknown.
Copyright © 2012 Elsevier Ltd. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Acyl Coenzyme A / chemistry
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Acyltransferases / chemistry*
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Acyltransferases / classification
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Acyltransferases / genetics
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Caffeic Acids / chemistry
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Chromatography, High Pressure Liquid
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Cloning, Molecular
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Coenzyme A-Transferases / chemistry
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Coenzyme A-Transferases / classification
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Coenzyme A-Transferases / genetics
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Coumaric Acids / chemistry*
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Enzyme Assays
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Escherichia coli / chemistry
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Escherichia coli / genetics
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Esterification
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Genes, Plant
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Glutathione Transferase / chemistry
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Glycerides / chemistry
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Glycerol / analogs & derivatives
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Glycerol / chemistry*
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Magnetic Resonance Spectroscopy
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Molecular Structure
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Oryza / chemistry
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Oryza / enzymology*
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Oryza / genetics
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Phylogeny
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Plant Proteins / chemistry
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Plant Proteins / genetics
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RNA, Plant / genetics
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / isolation & purification
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Shikimic Acid / chemistry
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Substrate Specificity
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Transformation, Genetic
Substances
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Acyl Coenzyme A
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Caffeic Acids
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Coumaric Acids
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Glycerides
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Plant Proteins
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RNA, Plant
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Recombinant Fusion Proteins
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Shikimic Acid
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ferulic acid
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Acyltransferases
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Glutathione Transferase
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Coenzyme A-Transferases
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Glycerol
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caffeic acid