Purification and characterization of a secretory lipolytic enzyme, MgLIP2, from Malassezia globosa.
Malassezia globosa is a lipid-dependent yeast that is found on the human skin and is associated with various skin disorders, including dandruff and seborrhoeic dermatitis (SD). Despite its important role in skin diseases, the molecular basis for its pathogenicity is poorly understood. The current hypothesis is that dandruff and SD are linked to fatty acid metabolism and secretory lipolytic enzymes, which hydrolyse sebaceous lipids and release irritating free fatty acids. A previous genomic analysis of M. globosa identified a family of 13 homologous genes predicted to encode secreted lipases. We have also reported that M. globosa had significantly higher extracellular lipase activity compared with other species. To identify the major secretory lipases of this yeast during its growth, we successfully purified and characterized an extracellular lipase MgLIP2. Based on MALDI-TOF MS, the peptide mass fingerprint of a tryptically digested protein MgLIP2 corresponded to ORF MGL_4054 of M. globosa. This lipase showed high esterase activity against 4-nitrophenyl palmitate and 1-naphthyl palmitate but not 1-naphthyl acetate. This enzyme had optimal activity at 30 °C and pH 5.0. Furthermore, the activity significantly increased in the presence of Triton X-100 and was partially inhibited by PMSF but was unaffected by univalent and divalent metal ions.
Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4295-B5 Nagatsuta, Midori-ku, Yokohama, Kanagawa 226-8501, Japan.
Weerapong
W
Juntachai
Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4295-B5 Nagatsuta, Midori-ku, Yokohama, Kanagawa 226-8501, Japan.
Takahiro
T
Oura
Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4295-B5 Nagatsuta, Midori-ku, Yokohama, Kanagawa 226-8501, Japan.
Susumu
S
Kajiwara
D003067:Coenzymes; D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D006801:Humans; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D008290:Malassezia / Q000201:enzymology* / Q000378:metabolism*; D008670:Metals; D009284:Naphthols / Q000378:metabolism; D017830:Octoxynol / Q000378:metabolism; D010168:Palmitates / Q000378:metabolism; D010664:Phenylmethylsulfonyl Fluoride / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity
D003067:Coenzymes; D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D008670:Metals; D009284:Naphthols; D010168:Palmitates; C050015:4-nitrophenyl palmitate; C005366:naphthyl acetate; D010664:Phenylmethylsulfonyl Fluoride; C018110:naphthyl palmitate; D017830:Octoxynol; D008049:Lipase