This project is directed towards a broad-based understanding of the manipulation of tissue cultures of Taxus brevifolia and related species for the production of taxol or taxol-like compounds which could be easily converted to taxol. Taxol is very promising anti-cancer agent which is in extremely short supply. A better understanding of how to manipulate cellular synthesis of taxol in tissue culture will provide the basis for efficient 'factory-like' production of taxol and thereby alleviate potential supply problems. The specific component goals of the project includes enabling methods such as development of rapid assay techniques for taxol; establishment of new cultures, particularly root cultures; evaluation of taxol self-inhibition and possible mechanisms of inhibition; and, characterization of taxol partition coefficients on a variety of resins and into organic solvents. With these techniques we will probe the effects of gas phase composition, elicitors, in situ product removal, and cell immobilization on cellular growth rates, taxol production rates, ratio of extracellular to intracellular taxol, and production of taxol-related compounds. Further, we will examine the effects of liquid shear in stirred reactors on cell physiology. Also, novel small-scale devices will be used to test the intrinsic capacity of root cultures for growth and product formation.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA055138-04
Application #
2096348
Study Section
Special Emphasis Panel (SRC (52))
Project Start
1991-08-15
Project End
1996-07-31
Budget Start
1994-08-01
Budget End
1996-07-31
Support Year
4
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Cornell University
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
City
Ithaca
State
NY
Country
United States
Zip Code
14850
Roberts, S C; Naill, M; Gibson, D M et al. (2003) A simple method for enhancing paclitaxel release from Taxus canadensis cell suspension cultures utilizing cell wall digesting enzymes. Plant Cell Rep 21:1217-20
Ketchum, R E; Tandon, M; Gibson, D M et al. (1999) Isolation of labeled 9-dihydrobaccatin III and related taxoids from cell cultures of taxuscell cultures of taxus canadensis elicited with m. J Nat Prod 62:1395-8
Ketchum, R E; Gibson, D M; Croteau, R B et al. (1999) The kinetics of taxoid accumulation in cell suspension cultures of Taxus following elicitation with methyl jasmonate. Biotechnol Bioeng 62:97-105
Hezari, M; Ketchum, R E; Gibson, D M et al. (1997) Taxol production and taxadiene synthase activity in Taxus canadensis cell suspension cultures. Arch Biochem Biophys 337:185-90
Mirjalili, N; Linden, J C (1996) Methyl jasmonate induced production of taxol in suspension cultures of Taxus cuspidata: ethylene interaction and induction models. Biotechnol Prog 12:110-8
Hefner, J; Rubenstein, S M; Ketchum, R E et al. (1996) Cytochrome P450-catalyzed hydroxylation of taxa-4(5),11(12)-diene to taxa-4(20),11(12)-dien-5alpha-ol: the first oxygenation step in taxol biosynthesis. Chem Biol 3:479-89
Srinivasan, V; Ciddi, V; Bringi, V et al. (1996) Metabolic inhibitors, elicitors, and precursors as tools for probing yield limitation in taxane production by Taxus chinensis cell cultures. Biotechnol Prog 12:457-65
Taticek, R A; Lee, C W; Shuler, M L (1994) Large-scale insect and plant cell culture. Curr Opin Biotechnol 5:165-74