A high-yield purification of glyoxalase i from rabbit liver by affinity chromatography on Blue Dextran-Sepharose 4B.
An improved method is described for the purification of glyoxalase I from rabbit liver. The method involves homogenization, ammonium sulfate precipitation followed by choloroform/ethanol precipitation, affinity chromatography on Blue Dextran-Sepharose 4B and chromatography on Sephadex G-100. The enzyme is specifically eluted from the affinity column by S-hexylglutathione, a competitive inhibitor of the enzyme. This procedure offers a convenient method for obtaining electrophoretically pure glyoxalase I in high yields (60--70%).
D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods; D003911:Dextrans; D007791:Lactoylglutathione Lyase / Q000302:isolation & purification*; D008099:Liver / Q000201:enzymology*; D008190:Lyases / Q000302:isolation & purification*; D008970:Molecular Weight; D011817:Rabbits; D012685:Sepharose / Q000031:analogs & derivatives
D003911:Dextrans; C024908:blue dextran-sepharose; D012685:Sepharose; D008190:Lyases; D007791:Lactoylglutathione Lyase