title: Cloning and expression in Escherichia coli of the gene for extracellular phospholipase A1 from Serratia liquefaciens.
abstract: From a genomic library of Serratia liquefaciens, a cloned DNA fragment comprising a two-gene operon was isolated and expressed in Escherichia coli. One of the gene products was identified as a phospholipase A1, and the enzyme was found to be excreted to the outer environment from S. liquefaciens as well as from E. coli. Both genes were sequenced, and the relationship between open reading frames in the DNA sequence and in vitro-expressed polypeptides was established. The length of the phospholipase polypeptide was found to be 319 amino acids. In the amino-terminal end of the coding sequence was a stretch of about 20 hydrophobic amino acids, but, in contrast to consensus signal peptides, no basic residues were present. The length of the second polypeptide was 227 amino acids. It was found that expression of the phospholipase gene in both E. coli and S. liquefaciens was growth phase regulated (late expression).
authors.affiliation: Department of Microbiology, Technical University of Denmark, Lyngby.
authors.forename: M
authors.initials: M
authors.lastname: Givskov
authors.forename: L
authors.initials: L
authors.lastname: Olsen
authors.forename: S
authors.initials: S
authors.lastname: Molin
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009876:Operon; D010740:Phospholipases / Q000235:genetics*; D010741:Phospholipases A / Q000096:biosynthesis / Q000235:genetics*; D054466:Phospholipases A1; D010957:Plasmids; D012705:Serratia / Q000201:enzymology / Q000235:genetics*
chemical_list: D010740:Phospholipases; D010741:Phospholipases A; D054466:Phospholipases A1