Cross-linked enzyme aggregates of pig liver esterase evaluated in kinetic resolution of racemic clopidogrel
Gaber, Yasser LU and Ismail, Mohamed LU
(2017)
In Biotechnology (Faisalabad)
16(3).
p.123-129
- Abstract
Background and Objective: Immobilization of enzymes as cross-linked aggregates is one of the cheapest, simplest and effective techniques for improving their stability and reusability and even avoiding contamination of the product with the catalyst. Clopidogrel is a widely used antiplatelet drug, only Sisomer is the biologically active enantiomer produced by resolution of the racemic compound. In the current study, cross-linked aggregates of pig liver esterase were prepared and evaluated for kinetic resolution of racemic clopidogrel. Materials and Methods: Cross-linked Enzyme Aggregates (CLEA) of the commercially available crude pig liver esterase cPLE were prepared using glutaraldehyde at concentrations of 12.5-125 mM as crosslinker... (More)
Background and Objective: Immobilization of enzymes as cross-linked aggregates is one of the cheapest, simplest and effective techniques for improving their stability and reusability and even avoiding contamination of the product with the catalyst. Clopidogrel is a widely used antiplatelet drug, only Sisomer is the biologically active enantiomer produced by resolution of the racemic compound. In the current study, cross-linked aggregates of pig liver esterase were prepared and evaluated for kinetic resolution of racemic clopidogrel. Materials and Methods: Cross-linked Enzyme Aggregates (CLEA) of the commercially available crude pig liver esterase cPLE were prepared using glutaraldehyde at concentrations of 12.5-125 mM as crosslinker either in presence or absence of Bovine serum albumin (BSA). cPLE-CLEA was used for kinetic resolution of racemic clopidogrel and compared to the performance of soluble cPLE. Light microscopy and scanning electron microscopy SEM were used to examine cPLE-CLEA. Results: Soluble cPLE showed ability to resolve racemic clopidogrel at enantioselectivity (E) of 9.2. The resolution of clopidogrel was found to be optimal at 30°C. The cPLE-CLEA preparations showed reduced enzymatic activity. The kinetic resolution experiments showed also lower E values (E = 1.3-4.5) compared to soluble cPLE. Microscopical examination of cPLE-CLEA showed wide size variation and SEM revealed the shape of cPLE-CLEA before and after use in the kinetic resolution experiments. Conclusion: Crude PLE was able to resolve racemic clopidogrel, the effects of different temperatures were studied and the highest E value recorded was 9.2 at 30°C. Increasing concentrations of glutaraldehyde as a cross-linker adversely affected PLE activity. The cPLE-CLEA showed lower enantioselectivity compared to the free cPLE.
(Less)
- author
- Gaber, Yasser
LU
and Ismail, Mohamed
LU
- organization
- publishing date
- 2017
- type
- Contribution to journal
- publication status
- published
- keywords
- CLEA, Clopidogrel, Enantioselectivity, Esterase, Glutaraldehyde, Kinetic resolution
- in
- Biotechnology (Faisalabad)
- volume
- 16
- issue
- 3
- pages
- 7 pages
- publisher
- Science Alert
- external identifiers
-
- scopus:85036477935
- ISSN
- 1682-296X
- DOI
- 10.3923/biotech.2017.123.129
- language
- English
- LU publication?
- yes
- id
- 5fbdf67f-e2c2-45ff-bc71-d04b419f3669
- date added to LUP
- 2023-08-25 12:34:11
- date last changed
- 2023-08-29 07:52:01
@article{5fbdf67f-e2c2-45ff-bc71-d04b419f3669,
abstract = {{<p>Background and Objective: Immobilization of enzymes as cross-linked aggregates is one of the cheapest, simplest and effective techniques for improving their stability and reusability and even avoiding contamination of the product with the catalyst. Clopidogrel is a widely used antiplatelet drug, only Sisomer is the biologically active enantiomer produced by resolution of the racemic compound. In the current study, cross-linked aggregates of pig liver esterase were prepared and evaluated for kinetic resolution of racemic clopidogrel. Materials and Methods: Cross-linked Enzyme Aggregates (CLEA) of the commercially available crude pig liver esterase cPLE were prepared using glutaraldehyde at concentrations of 12.5-125 mM as crosslinker either in presence or absence of Bovine serum albumin (BSA). cPLE-CLEA was used for kinetic resolution of racemic clopidogrel and compared to the performance of soluble cPLE. Light microscopy and scanning electron microscopy SEM were used to examine cPLE-CLEA. Results: Soluble cPLE showed ability to resolve racemic clopidogrel at enantioselectivity (E) of 9.2. The resolution of clopidogrel was found to be optimal at 30°C. The cPLE-CLEA preparations showed reduced enzymatic activity. The kinetic resolution experiments showed also lower E values (E = 1.3-4.5) compared to soluble cPLE. Microscopical examination of cPLE-CLEA showed wide size variation and SEM revealed the shape of cPLE-CLEA before and after use in the kinetic resolution experiments. Conclusion: Crude PLE was able to resolve racemic clopidogrel, the effects of different temperatures were studied and the highest E value recorded was 9.2 at 30°C. Increasing concentrations of glutaraldehyde as a cross-linker adversely affected PLE activity. The cPLE-CLEA showed lower enantioselectivity compared to the free cPLE.</p>}},
author = {{Gaber, Yasser and Ismail, Mohamed}},
issn = {{1682-296X}},
keywords = {{CLEA; Clopidogrel; Enantioselectivity; Esterase; Glutaraldehyde; Kinetic resolution}},
language = {{eng}},
number = {{3}},
pages = {{123--129}},
publisher = {{Science Alert}},
series = {{Biotechnology (Faisalabad)}},
title = {{Cross-linked enzyme aggregates of pig liver esterase evaluated in kinetic resolution of racemic clopidogrel}},
url = {{http://dx.doi.org/10.3923/biotech.2017.123.129}},
doi = {{10.3923/biotech.2017.123.129}},
volume = {{16}},
year = {{2017}},
}