Characterization of gamma-terpinene synthase from Citrus unshiu (Satsuma mandarin).
Gamma-terpinene is a monoterpene and a major component of essential oils made from citrus fruits and shows strong antioxidant activity in various assay systems. Plant gamma-terpinene synthase is a member of the monoterpene cyclase family, which produces a specific monoterpene through cyclization of geranyl diphosphate (GPP), but the monoterpene cyclases have not been fully characterized. It is necessary to prepare large amounts of gamma-terpinene synthase from Citrus unshiu (Satsuma mandarin) for the characterization, on this purpose we expressed the protein in Escherichia coli (E. coli) cells. As most monoterpene synthases have plastid-targeting signals, a gene lacking these signals was prepared and functionally expressed in E. coli cells harboring extra copies of the argU gene. The purified enzyme was incubated with GPP and the main product was confirmed to be gamma-terpinene by GC/MS.
Suzuki
Yukiko
Y
Department of Food and Nutritional Sciences, University of Shizuoka, 52-1 Yada, Shizuoka 422-8526, Japan.
Sakai
Hiroshi
H
Shimada
Takehiko
T
Omura
Mitsuo
M
Kumazawa
Shigenori
S
Nakayama
Tsutomu
T
D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism*; D002957:Citrus / Q000201:enzymology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology; D008401:Gas Chromatography-Mass Spectrometry; D010940:Plant Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism
D010940:Plant Proteins; D011994:Recombinant Proteins; D013729:Terpenes; D019883:Alkyl and Aryl Transferases; C022723:gamma-terpinene synthetase