title: Characterization of hydroxycinnamoyltransferase from rice and its application for biological synthesis of hydroxycinnamoyl glycerols.
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.
authors.affiliation: Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University, Seoul, Republic of Korea.
authors.forename: In A
authors.initials: IA
authors.lastname: Kim
authors.forename: Bong-Gyu
authors.initials: BG
authors.lastname: Kim
authors.forename: Mihyang
authors.initials: M
authors.lastname: Kim
authors.forename: Joong-Hoon
authors.initials: JH
authors.lastname: Ahn
mesh_terms: D000214:Acyl Coenzyme A / Q000737:chemistry; D000217:Acyltransferases / Q000737:chemistry* / Q000145:classification / Q000235:genetics; D002109:Caffeic Acids / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D015256:Coenzyme A-Transferases / Q000737:chemistry / Q000145:classification / Q000235:genetics; D003373:Coumaric Acids / Q000737:chemistry*; D057075:Enzyme Assays; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D004951:Esterification; D017343:Genes, Plant; D005982:Glutathione Transferase / Q000737:chemistry; D005989:Glycerides / Q000737:chemistry; D005990:Glycerol / Q000031:analogs & derivatives / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D012275:Oryza / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012765:Shikimic Acid / Q000737:chemistry; D013379:Substrate Specificity; D014170:Transformation, Genetic
chemical_list: D000214:Acyl Coenzyme A; D002109:Caffeic Acids; D003373:Coumaric Acids; D005989:Glycerides; D010940:Plant Proteins; D018749:RNA, Plant; D011993:Recombinant Fusion Proteins; D012765:Shikimic Acid; C004999:ferulic acid; D000217:Acyltransferases; D005982:Glutathione Transferase; D015256:Coenzyme A-Transferases; D005990:Glycerol; C040048:caffeic acid