Aqueous two-phase extraction of dehydrogenases using triazine dyes in PEG/phosphate systems

✍ Scribed by W.-H. Wang; R. Kuboi; I. Komasawa

• Publisher: Elsevier Science
• Year: 1992
• Tongue: English
• Weight: 891 KB
• Volume: 47
• Category: Article
• ISSN: 0009-2509
• DOI: 10.1016/0009-2509(92)80205-q

✦ Synopsis

AMract-Group-specific afEnity extraction of dehydrogenascs, such as bakerys past alcohol dchydrogenase, rabbit muscle lactic debydrogenase, bovine liver glutamic dehydrogenase and yeast gbuzose-6phosphate debydrogenase, was studied using triazine dyes in polyethylene glycol (PEG)/potassium pbospbate aqueous two-phase systems. The parameters and factors determining a%nity patition have heen evaluated. Triazine dye ligands were found to have approximately the same inhibition constants for alcohol and glutamic dehydrogenases in the presence of the top phase of PEG/phosphate syskm as those in the simple buffer solution without the two-phase system. Thus, tinity partitioning is observed for these enzymes in a PEG/phosphate system, except for a few enzymes wbicb have phosphate as an inhibitor or activator. PEG/phosphate systems have advantages of high ligand partition coefficients, high enzyme selectivities and low costs as compared to PEG/dcxtran systems. The selection and optimization of the affraity extraction processes have also been discussed.