Purification of phospholipase-C from Bacillus cereus by affinity chromatography on 2-(4-aminophenylsulphonyl)ethyl-cellulose.
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.
Kamberov
E
E
Department of Biochemistry and Microbiology, Plovdiv University, Bulgaria.
Ivanov
A
A
D000327:Adsorption; D001409:Bacillus cereus / Q000201:enzymology*; D001665:Binding Sites; D002482:Cellulose / Q000031:analogs & derivatives*; D002846:Chromatography, Affinity; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010767:Phosphorylcholine / Q000031:analogs & derivatives / Q000378:metabolism; D000077589:Sulfathiazole; D013432:Sulfathiazoles / Q000494:pharmacology; D014325:Tromethamine / Q000494:pharmacology; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism
C033523:4-nitrophenylphosphorylcholine; D013432:Sulfathiazoles; D014325:Tromethamine; D010767:Phosphorylcholine; C063737:2-(4-aminophenylsulfonyl)ethyl cellulose; D002482:Cellulose; D010738:Type C Phospholipases; D000077589:Sulfathiazole