Characterization of a neutral ceramidase orthologue from Aspergillus oryzae.
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 degrees C. The apparent K(m) and V(max) values of the enzyme for C12-NBD-ceramide were 3.32 microM and 0.085 micromol min(-1) mg(-1), respectively.
Tada
Sawaki
S
National Food Research Institute, Tsukuba, Ibaraki, Japan.
Matsushita-Morita
Mayumi
M
Suzuki
Satoshi
S
Kusumoto
Ken-Ichi
K
Kashiwagi
Yutaka
Y
D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000201:enzymology* / Q000235:genetics; D002518:Ceramides / Q000378:metabolism; D003001:Cloning, Molecular; D004795:Enzyme Stability; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D055576:Neutral Ceramidase / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D013696:Temperature
D002518:Ceramides; D011994:Recombinant Proteins; D055576:Neutral Ceramidase