Publication Type: | Journal Article |
Year of Publication: | 2004 |
Authors: | M. Hosoi, Ito, M., Yagura, T., Adams, R. Phillip, Honda, G. |
Journal: | Biol Pharm Bull |
Volume: | 27 |
Pagination: | 1979–85 |
Date Published: | Dec |
Keywords: | Amino Acid Sequence, DNA: Complementary, Gene Expression Regulation: Enzymologic, Intramolecular Lyases, Molecular Sequence Data, Perilla frutescens, Phylogeny |
Abstract: | 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. |