Molecular Cloning and Yeast Expression of Cinnamate 4-Hydroxylase from Ornithogalum saundersiae Baker
AbstractOSW-1, isolated from the bulbs of Ornithogalum saundersiae Baker, is a steroidal saponin endowed with considerable antitumor properties. Biosynthesis of the 4-methoxybenzoyl group on the disaccharide moiety of OSW-1 is known to take place biochemically via the phenylpropanoid biosynthetic pathway, but molecular biological characterization of the related genes has been insufficient. Cinnamic acid 4-hydroxylase (C4H, EC 22.214.171.124), catalyzing the hydroxylation of trans-cinnamic acid to p-coumaric acid, plays a key role in the ability of phenylpropanoid metabolism to channel carbon to produce the 4-methoxybenzoyl group on the disaccharide moiety of OSW-1. Molecular isolation and functional characterization of the C4H genes, therefore, is an important step for pathway characterization of 4-methoxybenzoyl group biosynthesis. In this study, a gene coding for C4H, designated as OsaC4H, was isolated according to the transcriptome sequencing results of Ornithogalum saundersiae. The full-length OsaC4H cDNA is 1,608-bp long, with a 1,518-bp open reading frame encoding a protein of 505 amino acids, a 55-bp 5′ non-coding region and a 35-bp 3'-untranslated region. OsaC4H was functionally characterized by expression in Saccharomyces cerevisiae and shown to catalyze the oxidation of trans-cinnamic acid to p-coumaric acid, which was identified by high performance liquid chromatography with diode array detection (HPLC-DAD), HPLC-MS and nuclear magnetic resonance (NMR) analysis. The identification of the OsaC4H gene was expected to open the way to clarification of the biosynthetic pathway of OSW-1.
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Kong, J.-Q.; Lu, D.; Wang, Z.-B. Molecular Cloning and Yeast Expression of Cinnamate 4-Hydroxylase from Ornithogalum saundersiae Baker. Molecules 2014, 19, 1608-1621.
Kong J-Q, Lu D, Wang Z-B. Molecular Cloning and Yeast Expression of Cinnamate 4-Hydroxylase from Ornithogalum saundersiae Baker. Molecules. 2014; 19(2):1608-1621.Chicago/Turabian Style
Kong, Jian-Qiang; Lu, Di; Wang, Zhi-Biao. 2014. "Molecular Cloning and Yeast Expression of Cinnamate 4-Hydroxylase from Ornithogalum saundersiae Baker." Molecules 19, no. 2: 1608-1621.