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Molecular characterization and sequence of phosphatidylinositol‐specific phospholipase C of Bacillus thuringiensis
Author(s) -
Lechner M.,
Kupke T.,
Stefanovic S.,
Götz F.
Publication year - 1989
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1989.tb00209.x
Subject(s) - biology , bacillus thuringiensis , biochemistry , escherichia coli , bacillus subtilis , peptide sequence , microbiology and biotechnology , phospholipase c , enzyme , gel electrophoresis , amino acid , molecular mass , polyacrylamide gel electrophoresis , recombinant dna , nucleic acid sequence , gene , bacteria , genetics
Summary 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, 38095). The NH 2 ‐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 34586. 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 sphingo‐myelin. The cleaving specifity of PI‐PLC was examined by thin layer chromatography.