title: Molecular cloning and expression of the gene encoding a phospholipase A1 from Aspergillus oryzae.
abstract: Phospholipase A1 (PLA1) is a hydrolytic enzyme that catalyzes removal of the acyl group from position 1 of lecithin to form lysolecithin. The genomic DNA and cDNA encoding PLA1 from Aspergillus oryzae were cloned with the mixed deoxyribonucleotide-primed polymerase chain reaction. The PLA1 gene is composed of 1,056 bp and has four exons and three short introns (63, 54, and 51 bp). The deduced amino acid sequence of PLA1 contained the N-terminal sequence of the mature PLA1 analyzed by Edman degradation. PLA1 cDNA has an open reading frame of 885 bp encoding the PLA1 precursor of 295 amino acid residues. The mature PLA1 is composed of 269 amino acid residues, and a prepro-sequence of 26 amino acid residues is at the N-terminal region of the PLA1 precursor. PLA1 has two possible N-glycosylation sites (Asn27 and Asn55). PLA1 has a consensus pentapeptide (-Gly-His-Ser-Xaa-Gly-), which is conserved in lipases. The amino acid sequence of PLA1 showed 47% identity with that of mono- and diacylglycerol lipase from Penicillium camembertii. The PLA1 cDNA was expressed in Saccharomyces cerevisiae KS58-2D, indicating the cloned gene to be functional.
authors.affiliation: Biomedical Research Laboratories, Sankyo Co., Ltd., Tokyo, Japan.
authors.forename: I
authors.initials: I
authors.lastname: Watanabe
authors.forename: R
authors.initials: R
authors.lastname: Koishi
authors.forename: Y
authors.initials: Y
authors.lastname: Yao
authors.forename: T
authors.initials: T
authors.lastname: Tsuji
authors.forename: N
authors.initials: N
authors.lastname: Serizawa
mesh_terms: D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000201:enzymology*; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010741:Phospholipases A / Q000235:genetics*; D054466:Phospholipases A1; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D010741:Phospholipases A; D054466:Phospholipases A1