1. Cloning and Characterization ofLonicera japonicap-Coumaroyl Ester 3-Hydroxylase Which Is Involved in the Biosynthesis of Chlorogenic Acid
- Author
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Peng Wang, Fengning Xiang, Zhenhua Liu, Gaobin Pu, and Bingqian Zhou
- Subjects
Models, Molecular ,DNA, Complementary ,Protein Conformation ,Ultraviolet Rays ,Molecular Sequence Data ,Sequence Homology ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,Biochemistry ,Japonica ,Mixed Function Oxygenases ,Analytical Chemistry ,chemistry.chemical_compound ,Plant Growth Regulators ,Chlorogenic acid ,Gene Expression Regulation, Plant ,Complementary DNA ,medicine ,Amino Acid Sequence ,Cloning, Molecular ,Molecular Biology ,Escherichia coli ,Southern blot ,chemistry.chemical_classification ,Methyl jasmonate ,biology ,Organic Chemistry ,General Medicine ,biology.organism_classification ,Molecular biology ,Lonicera ,Enzyme ,chemistry ,Heterologous expression ,Chlorogenic Acid ,Biotechnology - Abstract
Lonicera japonica is used in Chinese medicine as a source of antioxidants, primarily flavonoids, and a phenolic acid chlorogenic acid (CGA). Here we report the isolation and characterization of the full-length cDNA of LjC3H, a gene encoding p-coumaroyl ester 3-hydroxylase, an enzyme involved in CGA synthesis. Phylogenetic analysis indicated that is protein belongs to the CYP98A subfamily, and homology modeling revealed that its structure resembles that of other cytochrome P450 family proteins. Southern blot analysis indicated that more than one copy of sequences homologous to LjC3H is present in the L. japonica genome. Heterologous expression of LjC3H cDNA in Escherichia coli allowed an in vitro assay of LjC3H to be performed. This experiment revealed that the enzyme favors p-coumaroylshikimate over p-coumaroylquinate as substrate. LjC3H transcript abundance was increased both by treatment of the leaves with methyl jasmonate and by exposure to UV-B radiation. The CGA levels in the leaves of L. japonica were positively correlated with LjC3H transcript abundance.
- Published
- 2013