Open AccessIsolation, characterization and molecular cloning of a lipolytic enzyme secreted from Malassezia pachydermatis
Author(s) -
Shibata Nobuyuki,
Okanuma Noriko,
Hirai Kanako,
Arikawa Kumiko,
Kimura Mayumi,
Okawa Yoshio
Publication year - 2006
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2006.00106.x
Subject(s) - malassezia , isolation (microbiology) , molecular cloning , biology , enzyme , cloning (programming) , microbiology and biotechnology , biochemistry , gene , peptide sequence , computer science , programming language
Abstract Lipophilic Malassezia species may induce catheter‐associated sepsis in premature neonates and immunocompromised patients receiving parenteral lipid emulsions. To assess the participation of lipolytic enzymes in the pathogenesis of this yeast, we cloned a gene encoding the enzyme. A lipolytic enzyme in the culture supernatant of Malassezia pachydermatis was purified 210‐fold to homogeneity. The enzyme showed high esterase activity toward p ‐nitrophenyl octanoate. The cDNA encoding the enzyme was cloned using a degenerate oligonucleotide primer constructed from the N‐terminal amino acid sequence. The cDNA consisted of 1582 bp, including an open reading frame encoding 470 amino acids. The first 19 amino acids and the following 13 amino‐acid sequence were predicted to be the signal peptides for secretion and prosequence, respectively. The predicted molecular mass of the 438‐amino acid mature protein was 48 kDa. Analysis of the deduced amino acid sequence revealed that it contains the consensus motif (Gly–X–Ser–X–Gly), which is conserved among lipolytic enzymes. Homology investigations showed that the enzyme has similarities principally with 11 lipases produced by Candida albicans (29–34% identity) and some other yeast lipases.