Taxol biosynthesis: differential transformations of taxadien-5 alpha-ol and its acetate ester by cytochrome P450 hydroxylases from Taxus suspension cells.
The biosynthesis of the diterpenoid antineoplastic drug Taxol in Taxus species involves the cyclization of the ubiquitous isoprenoid intermediate geranylgeranyl diphosphate to taxa-4(5),11(12)-diene followed by cytochrome P450-mediated hydroxylation (with allylic rearrangement) of this olefin precursor to taxa-4(20),11(12)-dien-5 alpha-ol, and further oxygenation and acylation reactions. Based on the abundances of naturally occurring taxoids, the subsequent order of oxygenation of the taxane core is considered to occur at C10, then C2 and C9, followed by C13, and finally C7 and C1. Circumstantial evidence suggests that the acetylation of taxadien-5 alpha-ol may constitute the third specific step of Taxol biosynthesis. To determine whether taxadienol or the corresponding acetate ester serves as the direct precursor of subsequent oxygenation reactions, microsomal preparations isolated from induced Taxus cells and optimized for cytochrome P450 catalysis were incubated with each potential substrate. Both taxadienol and taxadienyl acetate were oxygenated to the level of a diol and to higher polyols at comparable rates by cytochrome P450 enzymes of the microsomal preparation. Preparative-scale incubation allowed the isolation of sufficient quantities of the diol derived from taxadienol to permit the NMR-based structural elucidation of this metabolite as taxa-4(20),11(12)-dien-5 alpha,13 alpha-diol, which may represent an alternate route of taxoid metabolism in induced cells. GC-MS-based structural definition of the diol monoacetate derived in microsomes from taxadienyl acetate confirmed this metabolite as taxa-4(20),11(12)-dien-5 alpha-acetoxy-10 beta-ol, thereby indicating that acetylation at C5 of taxadienol precedes the cytochrome P450-mediated insertion of the C10-beta-hydroxyl group of Taxol.
Wheeler
A L
AL
Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340, USA.
Long
R M
RM
Ketchum
R E
RE
Rithner
C D
CD
Williams
R M
RM
Croteau
R
R
D000215:Acylation; D001711:Biotransformation; D002474:Cell-Free System; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004952:Esters / Q000737:chemistry / Q000378:metabolism; D006900:Hydroxylation; D009682:Magnetic Resonance Spectroscopy; D008861:Microsomes / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000378:metabolism; D017239:Paclitaxel / Q000031:analogs & derivatives / Q000096:biosynthesis* / Q000378:metabolism*; D010936:Plant Extracts / Q000378:metabolism; D010946:Plants, Medicinal; D043823:Taxoids; D020946:Taxus / Q000378:metabolism*; D013997:Time Factors
D004952:Esters; D010936:Plant Extracts; D043823:Taxoids; C430093:taxa-4(20),11(12)-dien-5-yl acetate; C430092:taxa-4(20),11(12)-diene-5-ol; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D017239:Paclitaxel