A cDNA clone for 3-carene synthase from Salvia stenophylla.
The essential oil of Salvia stenophylla contains (+)-3-carene as the principal monoterpene component. Using an enriched cDNA library prepared from mRNA isolated from S. stenophylla peltate glandular trichomes, and a homology-based cloning strategy, a full-length cDNA was isolated that encoded a preprotein of 69.7 kDa which resembled a monoterpene synthase in sequence. Heterologous expression of the gene in Escherichia coli provided a soluble recombinant enzyme capable of catalyzing the divalent metal ion-dependent conversion of geranyl diphosphate to (+)-3-carene and to lesser amounts of limonene, myrcene, 4-carene and beta-phellandrene. This multiple-product synthase is responsible for the production of all of the essential oil monoterpenes of S. stenophylla.
Hoelscher
Dirk J
DJ
Institute of Biological Chemistry, Washington State University, PO Box 646340, Pullman, WA 99164-6340, USA.
Williams
David C
DC
Wildung
Mark R
MR
Croteau
Rodney
R
D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D015723:Gene Library; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000737:chemistry; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D027544:Salvia / Q000201:enzymology*; D012689:Sequence Homology, Nucleic Acid; D013729:Terpenes / Q000032:analysis
D018076:DNA, Complementary; D011994:Recombinant Proteins; D013729:Terpenes; D019883:Alkyl and Aryl Transferases