Glycosidation of betulinic acid using novozyme 435
In this study, 3-O-β-D-glucopyranoside-betulinic acid was successfully synthesized via the reaction between betulinic acid and glucose catalyzed by immobilized lipase from Candida antarctica (Novozyme 435) in t-butanol. The structure of the product obtained were elucidated using spectroscopic dat...
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| Format: | Thesis |
| Language: | English |
| Published: |
2016
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| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/69109/1/FS%202016%2037%20IR.pdf |
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| Summary: | In this study, 3-O-β-D-glucopyranoside-betulinic acid was successfully synthesized
via the reaction between betulinic acid and glucose catalyzed by immobilized lipase
from Candida antarctica (Novozyme 435) in t-butanol. The structure of the product
obtained were elucidated using spectroscopic data. Effects of individual reaction
parameters such as reaction time, reaction temperature, amount of enzyme and
substrate molar ratio, were investigated and optimized. The optimum conditions for
the reaction between betulinic acid and glucose were obtained at the reaction time of
48.50 h, temperature of 26oC, 175 mg of enzyme, and substrate molar ratio of 1:1.2;
giving 85.83 % of yield.
The Response Surface Methodology (RSM) based on five-level, three variables
Central Composite Rotatable Design (CCRD) was employed using Design Expert
software to evaluate the effect of synthesis parameters and its mutual interactions. It
was observed that the maximum conversion yield of 3-O-β-D-glucopyranosidebetulinic
acid 88.69% was obtained using 30.67 h, 54.30oC and 180 mg of enzyme
using betulinic acid (0.05 mmol) and glucose (0.1 mmol) respectively. The
experimental value obtained was 88.69%, closer to the results obtained using single
parameter.
Finally, the anticancer activity of the synthesized compound was evaluated against
cultured mouse embryonic fibroblast normal cell line (3T3), human cervical
carcinoma cancer (HeLa), human breast cancer (MCF-7), human T-promyelocytic
leukaemia (HL-60), and cell lines. From the results, BA showed high activity against
cultured human T-promyeloctic leukaemia (HL-60), human breast cancer (MCF-7),
and human cervical carcinoma cancer (HeLa) cell lines with IC50 values of MCF-7
0.8 μg/ml, HL-60 4.4 μg/ml and HeLa 4.8 μg/ml, respectively. On the other hand,
3-O-β-D-glucopyranoside-betulinic acid also showed strong activity against cultured,
HL-60, MCF-7and 3T3 with IC50 values of 8.4 μg/ml, 8.5 μg/ml and 2.75 μg/ml
respectively. However, it was found to have moderate activity against HeLa cell line
with IC50 value of 12.0 μg/ml.
In conclusion, an enzymatic synthesis of 3-O-β-D-glucopyranoside-betulinic acid
was successfully carried out by the reaction between betulinic acid and D-glucose in
an organic solvent using Novozyme 435. The activity of 3-O-β-D-glucopyranosidebetulinic
acid against cancer cell lines was found to be better than betulinic acid. |
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