| The essential oil-bearing plants produce a large variety of monoterpenes, the C;Garden sage (Salvia officinalis) produces a variety of monoterpenes, including enantiomeric forms of individual types. Two of the enzymes from sage which produce antipodal products, (+)-pinene cyclase and (;Many of the essential oil-bearing plants produce oxygenated derivatives of parent olefins via the intermediacy of cytochrome P-450 monoxygenases, such as sabinene hydroxylase from sage. In the process of synthesizing radiolabeled, photolabile inhibitors of the cytochrome P-450 monoterpene monoxygenases, a new method for tritium labeling of activated aromatic compounds was developed.;In an initial approach to the cloning and expression of cDNAs encoding monoterpene cyclases from sage, two very similar enzymes, (+)-bornyl pyrophosphate synthase and (+)-pinene synthase, were purified together, cleaved to peptides with CNBr, and the separated peptides subjected to N-terminal amino acid sequence analysis. This approach utilized a radiolabeled, mechanism-based inhibitor as both an aid to purification and to identify cyclase-derived peptides for sequencing. The peptide sequences were compared to the known sequence of (... |
