cDNA isolation and functional expression of myrcene synthase from Perilla frutescens.
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.
Hosoi
Madoka
M
Graduate School of Pharmaceutical Sciences, Kyoto University, Japan.
Ito
Michiho
M
Yagura
Toru
T
Adams
Robert Phillip
RP
Honda
Gisho
G
D000595:Amino Acid Sequence; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D019753:Intramolecular Lyases / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D031344:Perilla frutescens / Q000201:enzymology*; D010802:Phylogeny
D018076:DNA, Complementary; D019753:Intramolecular Lyases; C040265:pinene cyclase I