Cloning and structure of the mono- and diacylglycerol lipase-encoding gene from Penicillium camembertii U-150.
A gene (mdlA) encoding mono- and diacylglycerol lipase (MDGL) from Penicillium camembertii U-150 has been cloned using a 0.9-kb DNA fragment, generated by mixed oligodeoxyribonucleotide (oligo)-primed polymerase chain reaction (PCR), as a probe. Comparison of the nucleotide sequence of the gene and its cDNA clone, obtained by PCR, revealed the presence of two short introns (56 and 53 bp). Two transcription start points (tsp) were localized by primer extension analysis at 37 and 30 bp upstream from the ATG start codon and were preceded by the canonical TATAAA and CAAT sequences. The deduced amino acid (aa) sequence corresponds to 305 aa including a putative signal peptide of 26 aa. Despite significant differences in substrate specificity, the primary structure of the mature region shows homology (29% and 40%) to the triacylglycerol lipases from Mucor miehei and Humicola lanuginosa. Furthermore, the three residues presumed to form the catalytic site, serine, aspartic acid and histidine, are conserved. Primary structure comparisons of MDGL and triacylglycerol lipases are shown.
Yamaguchi
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S
Tsukuba Research Laboratories, Amano Pharmaceutical Co. Ltd., Ibaragi, Japan.
Mase
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Takeuchi
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K
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D003001:Cloning, Molecular; D005800:Genes, Fungal / Q000235:genetics*; D007438:Introns / Q000235:genetics; D008049:Lipase / Q000737:chemistry; D008071:Lipoprotein Lipase / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D010407:Penicillium / Q000201:enzymology / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D013329:Structure-Activity Relationship
D008049:Lipase; D008071:Lipoprotein Lipase