Synthesis of dye conjugates of ethylene oxide-propylene oxide copolymers and application in temperature-induced phase partitioning.
Synthesis of conjugates of the ethylene oxide/propylene oxide copolymer UCON 50-HB-5100 and the triazine dyes Cibacron Blue F3G-A and Procion Yellow HE-3G is described. The UCON-dye conjugate of Procion Yellow HE-3G is used as a ligand for affinity partitioning of glucose-6-phosphate dehydrogenase from bakers' yeast. The enzyme is first partitioned in a two-phase system composed of UCON, UCON-ligand and dextran, and the two phases isolated in separate containers. A small amount of salt is then added to the upper phase, which contains the UCON-ligand-enzyme complex, and the temperature increased above the cloud point of the UCON polymer to give a new two-phase system. The new two-phase system consists of an upper salt/water phase containing free enzyme and a lower UCON/water phase containing free UCON-ligand. Temperature-induced phase partitioning is thus seen to be of much assistance in dissociating enzyme-ligand complex, recovering enzyme and recycling UCON-ligand.
Alred
P A
PA
Department of Biochemistry, University of Lund, Sweden.
Tjerneld
F
F
Kozlowski
A
A
Harris
J M
JM
D055598:Chemical Phenomena; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002627:Chemistry, Physical; D004396:Coloring Agents / Q000138:chemical synthesis; D005656:Fungal Proteins / Q000302:isolation & purification; D005954:Glucosephosphate Dehydrogenase / Q000302:isolation & purification; D007700:Kinetics; D011095:Polyethylenes; D011126:Polypropylenes; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013696:Temperature; D014227:Triazines
D004396:Coloring Agents; D005656:Fungal Proteins; D011095:Polyethylenes; D011126:Polypropylenes; D014227:Triazines; C012842:Cibacron Blue F 3GA; C009794:procion yellow; C078661:UCON 50-HB-5100; D005954:Glucosephosphate Dehydrogenase