Biosynthesis of monoterpenes: stereochemistry of the coupled isomerization and cyclization of geranyl pyrophosphate to camphane and isocamphane monoterpenes.
The conversion of geranyl pyrophosphate to (+)-bornyl pyrophosphate and (+)-camphene is considered to proceed by the initial isomerization of the substrate to (-)-(3R)-linalyl pyrophosphate and the subsequent cyclization of this bound intermediate. In the case of (-)-bornyl pyrophosphate and (-)-camphene, isomerization of the substrate to the (+)-(3S)-linalyl intermediate precedes cyclization. The geranyl and linalyl precursors were shown to be mutually competitive substrates (inhibitors) of the relevant cyclization enzymes isolated from Salvia officinalis (sage) and Tanacetum vulgare (tansy) by the mixed substrate analysis method, demonstrating that isomerization and cyclization take place at the same active site. Incubation of partially purified enzyme preparations with (3R)-[1Z-3H]linalyl pyrophosphate plus [1-14C]geranyl pyrophosphate gave rise to double-labeled (+)-bornyl pyrophosphate and (+)-camphene, whereas incubation of enzyme preparations catalyzing the antipodal cyclizations with (3S)-[1Z-3H]-linalyl pyrophosphate plus [1-14C]geranyl pyrophosphate yielded double-labeled (-)-bornyl pyrophosphate and (-)-camphene. Each product was then transformed to the corresponding (+)- or (-)-camphor without change in the 3H:14C isotope ratio, and the location of the tritium label was deduced in each case by stereoselective, base-catalyzed exchange of the exo-alpha-hydrogen of the derived ketone. The finding that the 1Z-3H of the linalyl precursor was positioned at the endo-alpha-hydrogen of the corresponding camphor in all cases, coupled to the previously demonstrated retention of configuration at C1 of the geranyl substrate in these transformations, confirmed the syn-isomerization of geranyl pyrophosphate to linalyl pyrophosphate and the cyclization of the latter via the anti,endo- conformer. These relative stereochemical elements, in combination with the observed enantiospecificities of the enzymes for the linalyl intermediates, allows definition of the overall absolute stereochemistry of the coupled isomerization and cyclization of geranyl pyrophosphate to the antipodal camphane (bornane) and isocamphane monoterpenoids.
Croteau
R
R
Institute of Biological Chemistry, Washington State University, Pullman 99164-6340.
Gershenzon
J
J
Wheeler
C J
CJ
Satterwhite
D M
DM
D000080462:Acyclic Monoterpenes; D001892:Camphanes / Q000302:isolation & purification*; D002250:Carbon Radioisotopes; D055598:Chemical Phenomena; D002621:Chemistry; D006868:Hydrolysis; D007536:Isomerism; D039821:Monoterpenes; D010944:Plants / Q000201:enzymology*; D011106:Polyisoprenyl Phosphates / Q000096:biosynthesis / Q000378:metabolism*; D011865:Radioisotope Dilution Technique; D013729:Terpenes / Q000302:isolation & purification*; D014316:Tritium
D000080462:Acyclic Monoterpenes; D001892:Camphanes; D002250:Carbon Radioisotopes; D039821:Monoterpenes; D011106:Polyisoprenyl Phosphates; D013729:Terpenes; D014316:Tritium; C036751:linalyl pyrophosphate; C022082:bornyl pyrophosphate; C015234:geranyl pyrophosphate