Characterization of hydroxycinnamoyltransferase from rice and its application for biological synthesis of hydroxycinnamoyl glycerols
- 作者：In A Kim ; Bong-Gyu Kim ; Mihyang Kim ; Joong-Hoon Ahn ; jhahn@konkuk.ac.kr
- 关键词：Rice ; Oryza sativa ; Gramineae ; Hydroxycinnamic acid ; Hydroxycinnamic acid transferase
- 刊名：Phytochemistry
- 出版年：2012
- 期刊代码：85_00319422
- 类别：ch
- 出版时间：April, 2012
- 卷：76
- 期：Complete
- 页码：25-31
- 文件大小：560 K
摘要
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.