Purification, Immobilization and Characterization of Lipase Isoenzyme from Aspergillus niger with C8 Magnetic Particles

Samanta Hernández-García, María Inmaculada García-García, Francisco García-Carmona

Department of Biochemistry and Molecular Biology A, Faculty of Biology, Regional Campus of International Excellence “Campus Mare Nostrum”, University of Murcia, Campus Espinardo, Murcia, Spain.

DOI: 10.4236/abb.2014.57075                

Abstract

The purification of a lipase isoenzyme from an Aspergillus niger lipase A is reported in this manuscript. Purification was carried out in a simple adsorption step, in which the lipase was offered at low ionic strength to the commercially available C8 modified magnetic particles, MaKProt C8. When the isoenzyme was desorbed with a 0.2% solution of Triton X-100, the SDS-PAGE gel showed a single pure band with a molecular weight of 35 KDa. The purified fraction showed 66.75-fold purification compared with the crude extract. The pure fraction was characterized along with the crude extract and the lipase adsorbed on the MaKProt C8. The purified and the adsorbed lipase showed better activity for the tested substrates (p-nitrophenyl acetate, decanoate, myristate and palmitate) than the crude extract, the preferred substrates being myristate (26.7 μmol·min^-1·mg^-1) and decanoate (17.4 μmol·min^-1·mg^-1), respectively. The temperature and pH profiles showed no change for the three enzymes, the optimum temperature being 37°C and the best pH 7.0.

Keywords

Protein Purification, Protein Affinity, Magnetic Hydrophobic Particles, Protein Immobilization, Lipase Purification, Lipase Isoenzyme, C8 Magnetic Particles

Conflicts of Interest

The authors declare no conflicts of interest.

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