Optimized Synthesis of Lipase-Catalyzed 3-O-(3',3'-Dimethylsuccinyl)-Betulinic Acid by Immobilised Novozyme 435

The derivative of betulinic acid, 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (5) was successfully synthesized by the reaction of betulinic acid and 2,2-dimethylsuccinic anhydride, catalyzed by immobilized lipase from Candida antartica (Novozyme 435) in chloroform. The structure of the pro...

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Main Author:	Gunong @ Mohd Shah, Siti Aminah
Format:	Thesis
Language:	English
Published:	2010
Subjects:	Lipase - Case studies Acids - Analysis Enzymes - Analysis
Online Access:	http://psasir.upm.edu.my/id/eprint/19583/1/FS_2010_33_F.pdf
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id	my-upm-ir.19583
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spelling	my-upm-ir.195832015-05-19T02:47:32Z Optimized Synthesis of Lipase-Catalyzed 3-O-(3',3'-Dimethylsuccinyl)-Betulinic Acid by Immobilised Novozyme 435 2010-06 Gunong @ Mohd Shah, Siti Aminah The derivative of betulinic acid, 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (5) was successfully synthesized by the reaction of betulinic acid and 2,2-dimethylsuccinic anhydride, catalyzed by immobilized lipase from Candida antartica (Novozyme 435) in chloroform. The structure of the product was determined by spectroscopic methods. Effects of different reaction parameters were investigated and optimized in the model reaction. Optimum conditions to produce 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (5) up to 78.1 % were observed at reaction time; 24 h, amount of enzyme; 100 mg, betulinic acid (1) (0.055 mmole) to 2,2-dimethylsuccinic anhydride (0.055 mmole) substrate molar ratio; 1:1 at 50 °C. Response surface methodology (RSM) based on a five-level, three variables and central composite rotatable design (CCRD) was employed to evaluate the interactive effects of the parameters used in the synthesis methodology such as reaction time, temperature and enzyme amount. It was observed that, simultaneous increase in reaction time, temperature and amount of enzyme will increase the yields of 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (5). Based on the analysis of ridge max, the optimum conditions for the synthesis of 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (5) were as follows: 53.6 °C of reaction temperature, 28.15 hours of reaction time and 122 mg of enzyme for 1.0 mmol of betulinic acid (1) and 1.0 mmol of 2,2-dimethylsuccinic anhydride. The optimum predicted for percentage yield was at 83.93 % in which agree well with the actual value of 84.38 %. In brief, the anticancer activity of betulinic acid (1) and 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (5) were evaluated against cultured human T-promyelocytic leukemia (HL-60), human breast cancer (MCF-7), human cervical carcinoma cancer (HeLa) and mouse embryonic fibroblast normal cell line (3T3) cells lines. In particular, 3-O-(3',3'-dimethylsuccinyl)-betulinic acid showed nontoxic activity against human T-promyeloctic leukemia (HL-60) and human breast cancer (MCF-7) with IC50 > 30 μg/ml. However, it has better activity against human cervical carcinoma cancer (HeLa) (IC50 1.9 μg/ml) compared to betulinic acid (IC50 4.8 μg/ml). Interestingly, both compound were highly inactive against mouse embryonic fibroblast normal cell line (3T3) with IC50 > 30 μg/ml. Lipase - Case studies Acids - Analysis Enzymes - Analysis 2010-06 Thesis http://psasir.upm.edu.my/id/eprint/19583/ http://psasir.upm.edu.my/id/eprint/19583/1/FS_2010_33_F.pdf application/pdf en public masters Universiti Putra Malaysia Lipase - Case studies Acids - Analysis Enzymes - Analysis Faculty of Science
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English
topic	Lipase - Case studies Acids - Analysis Enzymes - Analysis
spellingShingle	Lipase - Case studies Acids - Analysis Enzymes - Analysis Gunong @ Mohd Shah, Siti Aminah Optimized Synthesis of Lipase-Catalyzed 3-O-(3',3'-Dimethylsuccinyl)-Betulinic Acid by Immobilised Novozyme 435
description	The derivative of betulinic acid, 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (5) was successfully synthesized by the reaction of betulinic acid and 2,2-dimethylsuccinic anhydride, catalyzed by immobilized lipase from Candida antartica (Novozyme 435) in chloroform. The structure of the product was determined by spectroscopic methods. Effects of different reaction parameters were investigated and optimized in the model reaction. Optimum conditions to produce 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (5) up to 78.1 % were observed at reaction time; 24 h, amount of enzyme; 100 mg, betulinic acid (1) (0.055 mmole) to 2,2-dimethylsuccinic anhydride (0.055 mmole) substrate molar ratio; 1:1 at 50 °C. Response surface methodology (RSM) based on a five-level, three variables and central composite rotatable design (CCRD) was employed to evaluate the interactive effects of the parameters used in the synthesis methodology such as reaction time, temperature and enzyme amount. It was observed that, simultaneous increase in reaction time, temperature and amount of enzyme will increase the yields of 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (5). Based on the analysis of ridge max, the optimum conditions for the synthesis of 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (5) were as follows: 53.6 °C of reaction temperature, 28.15 hours of reaction time and 122 mg of enzyme for 1.0 mmol of betulinic acid (1) and 1.0 mmol of 2,2-dimethylsuccinic anhydride. The optimum predicted for percentage yield was at 83.93 % in which agree well with the actual value of 84.38 %. In brief, the anticancer activity of betulinic acid (1) and 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (5) were evaluated against cultured human T-promyelocytic leukemia (HL-60), human breast cancer (MCF-7), human cervical carcinoma cancer (HeLa) and mouse embryonic fibroblast normal cell line (3T3) cells lines. In particular, 3-O-(3',3'-dimethylsuccinyl)-betulinic acid showed nontoxic activity against human T-promyeloctic leukemia (HL-60) and human breast cancer (MCF-7) with IC50 > 30 μg/ml. However, it has better activity against human cervical carcinoma cancer (HeLa) (IC50 1.9 μg/ml) compared to betulinic acid (IC50 4.8 μg/ml). Interestingly, both compound were highly inactive against mouse embryonic fibroblast normal cell line (3T3) with IC50 > 30 μg/ml.
format	Thesis
qualification_level	Master's degree
author	Gunong @ Mohd Shah, Siti Aminah
author_facet	Gunong @ Mohd Shah, Siti Aminah
author_sort	Gunong @ Mohd Shah, Siti Aminah
title	Optimized Synthesis of Lipase-Catalyzed 3-O-(3',3'-Dimethylsuccinyl)-Betulinic Acid by Immobilised Novozyme 435
title_short	Optimized Synthesis of Lipase-Catalyzed 3-O-(3',3'-Dimethylsuccinyl)-Betulinic Acid by Immobilised Novozyme 435
title_full	Optimized Synthesis of Lipase-Catalyzed 3-O-(3',3'-Dimethylsuccinyl)-Betulinic Acid by Immobilised Novozyme 435
title_fullStr	Optimized Synthesis of Lipase-Catalyzed 3-O-(3',3'-Dimethylsuccinyl)-Betulinic Acid by Immobilised Novozyme 435
title_full_unstemmed	Optimized Synthesis of Lipase-Catalyzed 3-O-(3',3'-Dimethylsuccinyl)-Betulinic Acid by Immobilised Novozyme 435
title_sort	optimized synthesis of lipase-catalyzed 3-o-(3',3'-dimethylsuccinyl)-betulinic acid by immobilised novozyme 435
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Science
publishDate	2010
url	http://psasir.upm.edu.my/id/eprint/19583/1/FS_2010_33_F.pdf
_version_	1747811417229099008

Optimized Synthesis of Lipase-Catalyzed 3-O-(3',3'-Dimethylsuccinyl)-Betulinic Acid by Immobilised Novozyme 435

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