Purification of The Lipase B of Candida antarctica from a Commercial Enzymatic Extract
Carlos R. Llerena-Suster, Laura E. Briand and Susana R. Morcelle
Affiliation: Centro de Investigación y Desarrollo en Ciencias Aplicadas-Dr. Jorge J. Ronco. Universidad Nacional de La Plata, CONICET, CCT La Plata. Calle 47 No 257, B1900AJK La Plata, Buenos Aires, Argentina.
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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