RESUMO
In this work, a novel electrochemical assay for characterizing both lipases and lipase inhibitors as well as for the determination of lipase activity is described. It is based on a carbon paste electrode, modified with cobalt(II)phthalocyanine, and multi-walled carbon nanotubes (MWCNTs). As reaction media, a sodium borate buffer was used (0.1 M, pH 9). The measurements were carried out in a batch system using differential pulse voltammetry (DPV) and 1,3-dilinolein as standard substrate. The activity assay showed a linearity for porcine pancreas lipase activity in a range between 20 and 300 U L-1 (per min) with a limit of detection (LOD) of 7 U L-1 and a limit of quantification (LOQ) of 20 U L-1. The kinetic behavior of the lipase reaction was investigated, resulting in a KM value of 0.29 mM. The applicability of the activity assay could be shown by investigating the activity of lipases from Aspergillus oryzae and Candida rugosa, and the results were confirmed by a reference method. The inhibitory effects were characterized with Orlistat.
