title: Molecular characterization and sequence of phosphatidylinositol-specific phospholipase C of Bacillus thuringiensis.
abstract: The gene encoding monophosphatidylinositol inositol phosphohydrolase (PI-specific phospholipase C, PI-PLC) of Bacillus thuringiensis was cloned in Staphylococcus carnosus TM300. The complete coding region comprises 987 base pairs corresponding to a precursor protein of 329 amino acids (molecular weight, 38,095). The NH2-terminal sequence of the purified enzyme from Escherichia coli indicated that the mature PI-PLC consists of 299 amino acid residues with a molecular weight of 34,586. Polyacrylamide gel electrophoresis revealed the same molecular weight for the purified enzyme isolated from the DNA-donor strain of B. thuringiensis and from the E. coli clone. By computer analysis, the secondary structure was predicted. The enzyme from the E. coli recombinant shows no activity on other phospholipids and sphingomyelin. The cleaving specificity of PI-PLC was examined by thin layer chromatography.
authors.affiliation: Institut für Biologie II, University of Tübingen, FRG.
authors.forename: M
authors.initials: M
authors.lastname: Lechner
authors.forename: T
authors.initials: T
authors.lastname: Kupke
authors.forename: S
authors.initials: S
authors.lastname: Stefanovic
authors.forename: F
authors.initials: F
authors.lastname: Götz
mesh_terms: D000595:Amino Acid Sequence; D001413:Bacillus thuringiensis / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D002855:Chromatography, Thin Layer; D003001:Cloning, Molecular; D003062:Codon; D004269:DNA, Bacterial; D003627:Data Interpretation, Statistical; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D054801:Phosphoinositide Phospholipase C; D010727:Phosphoric Diester Hydrolases / Q000235:genetics* / Q000302:isolation & purification; D010957:Plasmids; D011487:Protein Conformation; D013379:Substrate Specificity
chemical_list: D003062:Codon; D004269:DNA, Bacterial; D010446:Peptide Fragments; D010727:Phosphoric Diester Hydrolases; D054801:Phosphoinositide Phospholipase C; D043265:Phosphatidylinositol Diacylglycerol-Lyase