Cloning and nucleotide sequence of the mono- and diacylglycerol lipase gene ( mdlB) of Aspergillus oryzae

• Published in: FEMS microbiology letters Vol. 143; no. 1; pp. 63 - 67
• Main Authors: Tsuchiya, Atsushi, Nakazawa, Hidekazu, Toida, Jinichi, Ohnishi, Kunio, Sekiguchi, Junichi
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 15-09-1996
• Subjects: Aspergillus oryzae; Diacylglycerol; Lipase; mdlB gene; Monoacylglycerol; Nucleotide sequence; Aspergillus oryzae; Nucleotide sequence; mdlB gene; Lipase; Monoacylglycerol; Diacylglycerol
• DOI: 10.1016/0378-1097(96)00291-1
• EISSN: 1574-6968
• ISSN: 0378-1097

Aspergillus oryzae IFO4202 produces at least two extracellular lipolytic enzymes L1 and L2 (cutinase, and mono- and diacylglycerol lipase, respectively). Southern hybridization of restriction enzyme-digested genomic DNA fragments with 23mer oligonucleotides synthesized according to the amino acid sequence of the L2 as probe suggested the presence of the L2 gene (tentatively designated as
mdlB) and an additional weakly hybridizing region. A fragment containing the genomic
mdlB gene was cloned in
Escherichia coli. Nucleotide sequencing of the fragment revealed an open reading frame, comprising 1021 nucleotides, which contains two introns (51 and 52 nucleotides). Putative polyadenylation signals were found 182 and 287 bp downstream of the stop codon. The deduced amino acid sequence of the
mdlB gene corresponds to 306 amino acid residues including a leader sequence of 28 amino acids and is highly similar to that of the
mdlA gene of
Penicillium camembertii. Three residues presumed to form the catalytic triad (serine, aspartic acid and histidine) of lipases were also conserved.