Complete Purification of Phosphatidylinositol-Specific Phospholipase C from a Strain of Bacillus thuringiensis1

Abstract

A phosphatidylinositol-specific phospholipase C was purified from the culture broth of Bacillus thuringiensis IAM 12077 to a homogeneous state as revealed by polyacrylamide gel electrophoresis. The specific activity of the purified enzyme was 559 units/mg and recovery of the enzyme activity was 31%. Molecular and physiological properties of the purified enzyme, including molecular weight (22,000), isoelectric point (pI = 4.9) and its ectoenzyme-releasing activity, were studied in comparison with those other known enzymes of bacterial origin.

Direct evidence of involvement of glycosylphosphatidylinositol‐anchored proteins in the heavy metal‐mediated signal delivery into T lymphocytes

“…This transfectant was prepared by transfecting BW5147 Thy-l-a with PIG-A cDNA plasmid into which a neomycin resistance gene had been introduced, followed by selection in medium with G418 [23]. PIPLC was obtained from the culture supernatant of Bacillus thuringiensis, and was purified to give a single band on SDS-PAGE [24]. This PIPLC was protease-free and was not toxic to cells by the dye exclusion test at the concentration (500 mU/ml) used.…”

Release of Ectoenzymes from Small Intestine Brush Border Membranes of Mice by Phospholipases

“…PI-PLC was purified from the culture broth of B. thuringiensis lAM 12077 by the method of Ikezawa et al 15 ) Antiserum of a rabbit immunized by highly purified PI-PLe was prepared by the method reported previously. 9) Anti-PI-PLC IgG fraction was dialyzed against boratebuffered saline (pH 7.6).…”

Growth Inhibition, Morphological Change, and Ectoenzyme Release of LLC-PK1 Cells by Phosphatidylinositol-specific Phospholipase C ofBacillus thuringiensis

“…PI-PLC was purified from the culture broth of B. thuringiensis TAM 12077 by the method of Ikezawa et al 4 ) Antiserum from a rabbit immunized highly purified PI-PLC was prepared by a method reported previously. The anti-PI-PLC IgG fraction was dialyzed against boratebuffered saline (BBS.…”