Synthesis of Water Soluble Polymers with Covalently Bound General Ligands
The partition of biological macromolecules, cells, and cell organelles in aqueous polymer two-phase systems is a well known phenomenon (1). One of our research activities is concentrated on the design of liquid-liquid separation techniques based on this phenomenon for the isolation and purification of enzymes (2). The partition of the constituents of an enzyme extract in an aqueous two-phase system, consisting of polyethyleneglycol (PEG) and dextran or PEG and potassium phosphate, can be attributed mainly to nonspecific physico-chemical interactions. In principle the specificity might be enhanced by confining to one of the phases polymers with appropriate covalently bound ligands (affinity partition). Based on PEG, methods to synthesize water soluble polymers with covalently bound general ligands (NAD-, NADH-, AMP-, ATP- and pyridoxal-5-phosphate analogues) have been explored. The PEG-NAD(H) derivatives have been tested in affinity partition experiments with formate dehydrogenase and formaldehyde dehydrogenase. Their stability at different pH values and reutilization was investigated.
KeywordsPotassium Phosphate Enzymatic Oxidation Cell Organelle Water Soluble Polymer Biological Macromolecule
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