Affinity chromatography of lipoxygenases.
A number of aminohexyl agarose derivatives of unsaturated fatty acids have been prepared and evaluated as materials for the affinity chromatography of soybean and pea lipoxygenases. A practical method for a one-stage purification of soybean lipoxygenase-1, with a purification factor of 16, is described, using either linolenate or docosa-4,7,10,13,16,19-hexaenoate as ligands. Results show that alleged competitive inhibitors do not cause sharp elution from the affinity column, and that there is an increasing specificity of binding and sharpness of elution as the proportion of unsaturation in the ligand is increased. These results are discussed in terms of the relative importance of the types of bonding involved in enzyme-substrate binding.
D002846:Chromatography, Affinity; D005231:Fatty Acids, Unsaturated; D008041:Linoleic Acids; D008084:Lipoxygenase / Q000302:isolation & purification*; D010944:Plants / Q000201:enzymology; D011485:Protein Binding; D012685:Sepharose
D005231:Fatty Acids, Unsaturated; D008041:Linoleic Acids; D012685:Sepharose; D008084:Lipoxygenase