CLC number: S571.1
On-line Access: 2015-02-02
Received: 2014-07-08
Revision Accepted: 2014-10-23
Crosschecked: 2015-01-08
Cited: 1
Clicked: 5968
Citations: Bibtex RefMan EndNote GB/T7714
http://orcid.org/0000-0001-6738-6663
Yue Zhang, Hai-peng Lv, Cheng-ying Ma, Li Guo, Jun-feng Tan, Qun-hua Peng, Zhi Lin. Cloning of a caffeoyl-coenzyme A O-methyltransferase from Camellia sinensis and analysis of its catalytic activity[J]. Journal of Zhejiang University Science B, 2015, 16(2): 103-112.
@article{title="Cloning of a caffeoyl-coenzyme A O-methyltransferase from Camellia sinensis and analysis of its catalytic activity",
author="Yue Zhang, Hai-peng Lv, Cheng-ying Ma, Li Guo, Jun-feng Tan, Qun-hua Peng, Zhi Lin",
journal="Journal of Zhejiang University Science B",
volume="16",
number="2",
pages="103-112",
year="2015",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1400193"
}
%0 Journal Article
%T Cloning of a caffeoyl-coenzyme A O-methyltransferase from Camellia sinensis and analysis of its catalytic activity
%A Yue Zhang
%A Hai-peng Lv
%A Cheng-ying Ma
%A Li Guo
%A Jun-feng Tan
%A Qun-hua Peng
%A Zhi Lin
%J Journal of Zhejiang University SCIENCE B
%V 16
%N 2
%P 103-112
%@ 1673-1581
%D 2015
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1400193
TY - JOUR
T1 - Cloning of a caffeoyl-coenzyme A O-methyltransferase from Camellia sinensis and analysis of its catalytic activity
A1 - Yue Zhang
A1 - Hai-peng Lv
A1 - Cheng-ying Ma
A1 - Li Guo
A1 - Jun-feng Tan
A1 - Qun-hua Peng
A1 - Zhi Lin
J0 - Journal of Zhejiang University Science B
VL - 16
IS - 2
SP - 103
EP - 112
%@ 1673-1581
Y1 - 2015
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1400193
Abstract: Epigallocatechin-3-O-(3-O-methyl) gallate (EGCG3"Me) present in leaves of Camellia sinensis has many beneficial biological activities for human health. However, EGCG3"Me occurs naturally in tea leaves in extremely limited quantities. Finding an enzyme from C. sinensis to catalyze the synthesis of EGCG3"Me is an alternative method to make up for the scarcity of EGCG3"Me in natural situations. In the present study, a complementary DNA (cDNA) encoding region and genomic DNA of the caffeoyl-coenzyme A O-methyltransferase (CCoAOMT) gene were isolated from C. sinensis (designated CsCCoAOMT). Nucleotide sequence analysis of CsCCoAOMT revealed an open reading frame of 738 bp that encodes a polypeptide with a predicted molecular weight of 28 kDa, which correlated well with the results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The full-length DNA sequence (2678 bp) contained five exons and four introns. The deduced amino acid sequence of CsCCoAOMT shared 92% identity with CCoAOMTs from Codonopsis lanceolata and Betula luminifera. The catalytic activity of CsCCoAOMT was analyzed. Three monomethylated epigallocatechin-3-O-gallate (EGCG) compounds (EGCG4"Me, EGCG3"Me, and EGCG3'Me) were produced by CsCCoAOMT with Km in the micromolar range. Real-time polymerase chain reaction (RT-PCR) experiments indicated that the CsCCoAOMT transcript was present at low levels during the early stages of leaf maturity (the first leaf and bud on a shoot) but the relative expression was augmented at advanced stages of leaf maturity (the third or fourth leaf on a shoot), which accorded well with changes in EGCG3"Me content in fresh leaves. Hence, we concluded that CsCCoAOMT catalyzes the syntheses of methylated EGCGs.
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