Abstract
This paper presents a rational strategy to purify the lipase B of Candida antarctica of the commercial extract Lipozyme® through size exclusion coupled with anionic exchange chromatography using a non conventional, easy to remove buffer system such as ammonia-ammonium. In this context, each step of the purification was followed through the determination of the protein content, esterase activity measurements, SDS-PAGE, agarose electrophoresis, UVspectroscopy and isoelectric focusing. The purification of the commercial extract afforded a sample that retains 47% of the proteins (being CALB the major enzymatic component of the purified sample) with a hydrolytic activity higher than the starting crude extract.
Keywords: Protein purification, CALB, UV absorbance, Bradford, size exclusion chromatography, Ion exchange chromatography.
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Current Catalysis
Title:Purification of The Lipase B of Candida antarctica from a Commercial Enzymatic Extract
Volume: 3
Author(s): Carlos R. Llerena-Suster, Laura E. Briand and Susana R. Morcelle
Affiliation:
Keywords: Protein purification, CALB, UV absorbance, Bradford, size exclusion chromatography, Ion exchange chromatography.
Abstract: This paper presents a rational strategy to purify the lipase B of Candida antarctica of the commercial extract Lipozyme® through size exclusion coupled with anionic exchange chromatography using a non conventional, easy to remove buffer system such as ammonia-ammonium. In this context, each step of the purification was followed through the determination of the protein content, esterase activity measurements, SDS-PAGE, agarose electrophoresis, UVspectroscopy and isoelectric focusing. The purification of the commercial extract afforded a sample that retains 47% of the proteins (being CALB the major enzymatic component of the purified sample) with a hydrolytic activity higher than the starting crude extract.
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Cite this article as:
Llerena-Suster R. Carlos, Briand E. Laura and Morcelle R. Susana, Purification of The Lipase B of Candida antarctica from a Commercial Enzymatic Extract, Current Catalysis 2014; 3 (2) . https://dx.doi.org/10.2174/2211544702666131224234223
DOI https://dx.doi.org/10.2174/2211544702666131224234223 |
Print ISSN 2211-5447 |
Publisher Name Bentham Science Publisher |
Online ISSN 2211-5455 |
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