Partial purification and characterization of two sesquiterpene cyclases from sage (Salvia officinalis) which catalyze the respective conversion of farnesyl pyrophosphate to humulene and caryophyllene.
Humulene cyclase and caryophyllene cyclase, two enzymes which catalyze the cyclization of farnesyl pyrophosphate to the respective sesquiterpene olefins, have been partially purified from the supernatant fraction of a sage (Salvia officinalis) leaf epidermis extract and separated from each other by a combination of hydrophobic interaction, gel filtration, and ion-exchange chromatography. The molecular weight of both cyclases was estimated by gel filtration to be 57,000 and both cyclases exhibited a pH optimum of 6.5 and preferred Mg2+ (Km approximately 1.5 mM) as the required divalent metal cation. Both enzymes possessed a Km of about 1.7 microM for farnesyl pyrophosphate, were strongly inhibited by p-hydroxymercuribenzoate, and exhibited comparable sensitivities to a variety of other potential inhibitors. The properties of the two sesquiterpene olefin cyclases, which are the first from a higher plant source to be examined in detail, were very similar to each other and to other monoterpene, sesquiterpene, and diterpene cyclases previously described.
Dehal
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Institute of Biological Chemistry, Washington State University, Pullman 99164-6340.
Croteau
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D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003500:Cyclization; D007535:Isomerases; D000081222:Monocyclic Sesquiterpenes; D010944:Plants / Q000201:enzymology*; D000081223:Polycyclic Sesquiterpenes; D011106:Polyisoprenyl Phosphates / Q000378:metabolism*; D012717:Sesquiterpenes / Q000378:metabolism*; D014166:Transferases / Q000302:isolation & purification*
D000081222:Monocyclic Sesquiterpenes; D000081223:Polycyclic Sesquiterpenes; D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; C042686:humulene; C004808:farnesyl pyrophosphate; C024714:caryophyllene; D014166:Transferases; D007535:Isomerases; C055295:caryophyllene cyclase; C055294:humulene cyclase