A single-step large-scale purification of pyruvate oxidase.
Pyruvate oxidase is an Escherichia coli peripheral membrane flavoprotein which catalyzes the oxidative decarboxylation of pyruvate to acetate and CO2. Pyruvate oxidase, like several other peripheral membrane enzymes, can be activated either by binding to lipid amphiphiles or by limited protease digestion. This paper reports a rapid and convenient method for effecting the large-scale purification of pyruvate oxidase from crude enzyme preparations using a Triton X-114 phase separation technique. It appears likely that this purification procedure can be used successfully with the family of enzymes which respond to both lipid and protease activation.
Zhang
T F
TF
Roger Adams Laboratory, Department of Biochemistry, University of Illinois, Urbana 61801.
Hager
L P
LP
D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology*; D007202:Indicators and Reagents; D008970:Molecular Weight; D017830:Octoxynol; D011092:Polyethylene Glycols; D011771:Pyruvate Oxidase / Q000302:isolation & purification*; D014461:Ultracentrifugation / Q000379:methods
D003902:Detergents; D007202:Indicators and Reagents; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; D011771:Pyruvate Oxidase