Preparation of amine oxidase from bovine serum by affinity chromatography on aminoheyxyl-Sepharose.
Amine oxidase was purified from bovine serum by affinity chromatography on aminohexyl substituted Sepharose. The enzyme was adsorbed on the chromatographic support in a suspension of aminohexyl Sepharose in diluted serum. After thorough washing with buffer, the gel was packed in a column and the enzyme eluted with 10 mM octylamine. Using this procedure it was possible to obtain apparently homogeneous amine oxidase in a single-step procedure. The specific enzyme activity was 0.14 mumoles benzaldehyde formed per minute at 25 degrees C per mg enzyme protein. Based on the activity of amine oxidase in serum, the yield of enzyme was 64%.
D006631:Amine Oxidase (Copper-Containing); D000588:Amines / Q000097:blood / Q000302:isolation & purification; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000097:blood / Q000302:isolation & purification*; D012685:Sepharose / Q000031:analogs & derivatives
D000588:Amines; C024903:aminohexyl-sepharose; D012685:Sepharose; D006631:Amine Oxidase (Copper-Containing); D000587:Oxidoreductases Acting on CH-NH Group Donors