Characterization of a neutral ceramidase orthologue from Aspergillus oryzae

Abstract

Ceramide is an important molecule not only structurally but also regulationally as a modulator of various cellular events. Ceramidase (CDase) are classified into three different types (acid, alkaline, and neutral CDases). Neutral CDase could play an important role in the regulation of ceramide levels in the extracellular space. In this study, we describe the characterization of a neutral CDase orthologue from the filamentous fungus Aspergillus oryzae. The gene encoding the neutral CDase orthologue was cloned and overexpressed in A. oryzae. The purified recombinant enzyme was optimally active at pH 4.0-4.5 and 40°C. The apparent Km and Vmax values of the enzyme for C12-NBD-ceramide were 3.32 μM and 0.085 μmol min-1 mg -1, respectively. © 2009 National Food Research.