Phospholipase C from Clostridium perfringens: preparation and characterization of homogeneous enzyme.

Abstract:

:A new procedure for the purification of phospholipase C from Clostridium perfringens has been devised that results in essentially pure enzyme. The procedure consists of ammonium sulfate fractionation, ion-exchange chromatography on QAE-Sephadex, and affinity chromatography on phosphatidylcholine linked to Sepharose. The molecular weight of the enzyme, determined by sodium dodecyl sulfate-gel electrophoresis, amino acid analysis, and gel filtration, is 43,000; and the isoelectric point is pH 5.4. The enzyme was optimally active with phosphatidylcholine dispersed in sodium deoxycholate, although appreciable activity was observed with either phosphatidylcholine or sphingomyelin dispersed with ethanol. The requirement for metal ions in the assay could be met by a number of different ions. The pure enzyme was found to contain 2 mol zinc per mol enzyme, thus implicating it as a zinc metalloenzyme.

journal_name

Arch Biochem Biophys

authors

Krug EL,Kent C

doi

10.1016/0003-9861(84)90403-x

subject

Has Abstract

pub_date

1984-06-01 00:00:00

pages

400-10

issue

2

eissn

0003-9861

issn

1096-0384

pii

0003-9861(84)90403-X

journal_volume

231

pub_type

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