cDNA isolation and functional expression of myrcene synthase from Perilla frutescens

Publication Type:Journal Article
Year of Publication:2004
Authors:M. Hosoi, Ito, M., Yagura, T., Adams, R. Phillip, Honda, G.
Journal:Biol Pharm Bull
Date Published:Dec
Keywords:Amino Acid Sequence, DNA: Complementary, Gene Expression Regulation: Enzymologic, Intramolecular Lyases, Molecular Sequence Data, Perilla frutescens, Phylogeny

cDNA cloning of a monoterpene synthase from Perilla frutescens whose steam-distilled oil contains 92.9% perillaketone, was performed by the PCR method using primers designed based on limonene synthase. The full-length nucleotide sequence of this cDNA consisted of 1978 bp including a 1827-bp translational region encoding a deduced protein of 608 amino acids, which was similar to that of limonene synthase from P. frutescens (85% identity). Functional expression of this clone in Escherichia coli yielded an active monoterpene synthase enzyme, which converted geranyl diphosphate into 53.8% myrcene, 20.9% sabinene, 19.8% linalool and 5.5% limonene. As for the extraction of reaction products, we performed SPME (solid phase micro extraction) as well as conventional solvent extraction, and compared these two extraction methods.

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