Selective separation process of proteins based on the heat stress-induced translocation across phospholipid membranes.
Heating of several protein solutions at 40-47 degrees C for 5-60 min in the presence of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) liposomes induced the translocation of beta-galactosidase (beta-gal), alpha-glucosidase (alpha-glu) and bovine carbonic anhydrase (CAB) from outer to inner aqueous phase across the liposome membrane. The translocated amounts of beta-gal at various temperatures were maximized under suitable heating conditions (45 degrees C, 30 min). Those of alpha-glu and CAB were maximized at 40-45 and 60 degrees C, respectively. Each maximum value could be correlated with the corresponding local hydrophobicity of each protein evaluated by the aqueous two-phase partitioning method. The possibility to apply these heat-induced translocation phenomena to the bioseparation of proteins was successfully demonstrated for the model mixture solution of beta-gal, alpha-glu and CAB.
Department of Chemical Science and Engineering, Graduate School of Engineering Science, Osaka University, Toyonaka, Japan.
H
H
Umakoshi
T
T
Shimanouchi
R
R
Kuboi
D001692:Biological Transport; D002256:Carbonic Anhydrases / Q000737:chemistry; D002623:Chemistry Techniques, Analytical / Q000379:methods; D006358:Hot Temperature; D008081:Liposomes; D010713:Phosphatidylcholines; D000520:alpha-Glucosidases / Q000737:chemistry; D001616:beta-Galactosidase / Q000302:isolation & purification*
D008081:Liposomes; D010713:Phosphatidylcholines; D000520:alpha-Glucosidases; D001616:beta-Galactosidase; D002256:Carbonic Anhydrases; C028694:1-palmitoyl-2-oleoylphosphatidylcholine