Abstract
Objectives
To clone and characterize caffeic acid 3-O-methyltransferase (LcCOMT) from the rhizome of Ligusticum chuanxiong, a traditional medicinal herb having a high content of ferulic acid.
Results
LcCOMT encoded an ORF of 362 amino acids with a calculated MW of 39,935 Da and pI of 5.94. Polygenetic tree indicated that LcCOMT was attributed to a new member of COMTs in plants. The recombinant LcCOMT was expressed in E. coli. HPLC and 1H NMR analyses of purified LcCOMT protein confirmed that it could catalyze caffeic acid to produce ferulic acid in vitro. The further site-mutagenesis proved that His268 was one key catalytic residue. In addition, the substantial changing expression level of LcCOMT under chilling treatment suggested that LcCOMT might play important role in the accumulation of ferulic acid under chilling treatment.
Conclusions
This is the first report of the isolation and characterization of a COMT clone from traditional medicine containing high contents of pharmaceutical ferulic acid.
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Acknowledgments
This work was supported by the Grant (No. 31371232) from National Natural Science Foundation of China, Grant (2014ZX09304307001-019) from National Science and Technology Major Project and Grant (No. 2682014RC14) from Fundamental Research Funds for the Central Universities of China.
Supporting information
Supplementary Fig. 1—Sequence alignment of LcCOMT with other SAM-dependent OMT family members.
Supplementary Fig. 2—Phylogenetic tree of LcCOMT and other OMTs constructed by neighbor-joining algorithm.
Supplementary Fig. 3—1H NMR chemical shifts (ppm) for ferulic acid.
Supplementary Table 1—Comparison of the deduced amino acid sequences between LcCOMT and known SAM-dependent O-methyltransferases.
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Li, JJ., Zhang, G., Yu, Jh. et al. Molecular cloning and characterization of caffeic acid 3-O-methyltransferase from the rhizome of Ligusticum chuanxiong . Biotechnol Lett 37, 2295–2302 (2015). https://doi.org/10.1007/s10529-015-1917-y
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DOI: https://doi.org/10.1007/s10529-015-1917-y