Purification of phospholipase-C from Bacillus cereus by affinity chromatography on 2-(4-aminophenylsulphonyl)ethyl-cellulose

J Chromatogr. 1990 Feb 23;525(2):307-18. doi: 10.1016/s0378-4347(00)83407-6.

Abstract

A new method for the purification of phospholipase-C (phosphatidylcholine cholinephosphohydrolase, EC 3.1.4.3) from Bacillus cereus has been developed, based on its affinity to 2-(4-aminophenylsulphonyl)ethyl derivative of beaded cellulose. The enzyme was adsorbed on the affinity sorbent through a site(s) that was clearly distinct from its catalytically active site, because it was still active in the immobilized state. A possible role of enzyme-inhibitor interaction in enzyme binding to the ligand used is discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adsorption
  • Bacillus cereus / enzymology*
  • Binding Sites
  • Cellulose / analogs & derivatives*
  • Chromatography, Affinity*
  • Hydrogen-Ion Concentration
  • Kinetics
  • Phosphorylcholine / analogs & derivatives
  • Phosphorylcholine / metabolism
  • Sulfathiazole
  • Sulfathiazoles / pharmacology
  • Tromethamine / pharmacology
  • Type C Phospholipases / antagonists & inhibitors
  • Type C Phospholipases / isolation & purification*
  • Type C Phospholipases / metabolism

Substances

  • 4-nitrophenylphosphorylcholine
  • Sulfathiazoles
  • Tromethamine
  • Phosphorylcholine
  • 2-(4-aminophenylsulfonyl)ethyl cellulose
  • Cellulose
  • Type C Phospholipases
  • Sulfathiazole