Effect of polymer structure on affinity partitioning of lactate dehydrogenase in polymer-water two-phase systems.

Lactate dehydrogenase from rabbit muscle (EC 1.1.1.27) was partitioned between the liquid phases of aqueous two-phase systems containing two kinds of polymers. The partition was influenced by the use of a dye ligand, Procion Yellow HE-3G, bound to a polymer. The ligand-carrying polymers were poly(ethyleneglycol), poly(vinyl alcohol), ethylhydroxyethylcellulose, hydroxypropyl-starch, Ficoll and dextran. Their influence on the partition of lactate dehydrogenase was studied in systems composed of the following pairs of polymers: dextran-poly(ethylene glycol), dextran-poly(vinyl alcohol), dextran-Ficoll and hydroxypropyl-starch-poly (ethylene glycol). The ratio between the change in the logarithmic partition coefficient of the enzyme, delta log K, caused by the ligand and the logarithm of the partition coefficient of the ligand-polymer, log KL-polymer, was used as a measure of the number of ligand polymers bound per enzyme molecule under saturation conditions. Possible explanations for the observed effects are discussed.