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Isolation and identification of secondary metabolites from taxus media suspension cell culture Pawlak, Sonia Kristin
Abstract
The diterpenoid taxol, a compound isolated from the bark of the Pacific Yew tree, Taxus brevefolia, has shown remarkable success in the treatment of several types of cancer. Limited supply of taxol, due to low concentrations produced naturally and difficulty in synthesis of the drug, have spurred research attempts to develop alternative methods of producing taxane compounds in vitro using suspension cell cultures. Suspension cell culture propagated from the stems of Taxus media were grown in airlift type bioreactors for 28 days using a modified B5 medium. The cells and broth from multiple bioreactor experiments were combined and extracted with ethyl acetate and methanol. The ethyl acetate extracts were roughly separated by partitioning with solvents before high pressure liquid chromatography (HPLC) analysis was performed to determine if taxol, baccatin III, 10 deacetylbaccatin, 14-hydroxy-10-deacetylbaccatin, 14- hydroxybaccatin, cephlomannine, 7-epitaxol or 7-methyltaxol were present. Additionally, the partitioned extract was checked agains the taxane standards by thin layer chromatography (TLC). There was no indication that the taxane compounds used as standards were present in the cell extract above the detectable limits for these methods. To determine what types of compounds the cell culture was producing, the cell extract was separated using column chromatography, preparative TLC and recrystallization. Five compounds were purified and identified as 3-β-0[P- β -glucoside] β -sitosterol, 3B-6B, 23-trihydroxyolean-28-oic acid, cyclo (L-proline-L- valine), by comparing mass spectral (MS) and nuclear magnetic resonance (NMR) spectroscopy with that published in the literature. Two novel compounds 9,10 epoxy, 11-hydroxy 12,13 linoleic acid and 4,6- dihydroxy,-2-methoxy, 8-methyl xanthone were found in cell culture. They were identified using NMR and MS data and by comparison with compounds of like structure in the literature.
Item Metadata
Title |
Isolation and identification of secondary metabolites from taxus media suspension cell culture
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Creator | |
Publisher |
University of British Columbia
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Date Issued |
1999
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Description |
The diterpenoid taxol, a compound isolated from the bark of the Pacific Yew tree,
Taxus brevefolia, has shown remarkable success in the treatment of several types of
cancer. Limited supply of taxol, due to low concentrations produced naturally and
difficulty in synthesis of the drug, have spurred research attempts to develop alternative
methods of producing taxane compounds in vitro using suspension cell cultures.
Suspension cell culture propagated from the stems of Taxus media were grown in
airlift type bioreactors for 28 days using a modified B5 medium. The cells and broth from
multiple bioreactor experiments were combined and extracted with ethyl acetate and
methanol. The ethyl acetate extracts were roughly separated by partitioning with solvents
before high pressure liquid chromatography (HPLC) analysis was performed to determine
if taxol, baccatin III, 10 deacetylbaccatin, 14-hydroxy-10-deacetylbaccatin, 14-
hydroxybaccatin, cephlomannine, 7-epitaxol or 7-methyltaxol were present.
Additionally, the partitioned extract was checked agains the taxane standards by thin
layer chromatography (TLC). There was no indication that the taxane compounds used
as standards were present in the cell extract above the detectable limits for these methods.
To determine what types of compounds the cell culture was producing, the cell
extract was separated using column chromatography, preparative TLC and
recrystallization. Five compounds were purified and identified as 3-β-0[P- β -glucoside]
β -sitosterol, 3B-6B, 23-trihydroxyolean-28-oic acid, cyclo (L-proline-L- valine), by
comparing mass spectral (MS) and nuclear magnetic resonance (NMR) spectroscopy
with that published in the literature. Two novel compounds 9,10 epoxy, 11-hydroxy
12,13 linoleic acid and 4,6- dihydroxy,-2-methoxy, 8-methyl xanthone were found in
cell culture. They were identified using NMR and MS data and by comparison with
compounds of like structure in the literature.
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Extent |
5355517 bytes
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Genre | |
Type | |
File Format |
application/pdf
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Language |
eng
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Date Available |
2009-06-26
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Provider |
Vancouver : University of British Columbia Library
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Rights |
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
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DOI |
10.14288/1.0228823
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URI | |
Degree | |
Program | |
Affiliation | |
Degree Grantor |
University of British Columbia
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Graduation Date |
1999-11
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Campus | |
Scholarly Level |
Graduate
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Aggregated Source Repository |
DSpace
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Item Media
Item Citations and Data
Rights
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.