Abstract
Textile dyes have been used in polymer-bound state for selective extraction of enzymes within aqueous two-phase systems composed of two polymers. The effectivity of this extraction depends on several factors including type, molecular weight and concentration of bulk polymers as well as the ligand-carrying polymer. Also the degree of substitution (mole ligand/mole polymer) is a parameter which now has been studied. A high degree of substitution does not only give rise to affinity partition but also allows steering of the partition of the ligand-polymer itself. Therefore, the same ligand-polymer can be used for extraction of enzymes both to the upper and to lower phase. Studies have been carried out with Procion yellow HE-3G bound to poly(ethylene glycol), dextran, hydroxypropyl starch, Ficoll, ethylhydroxyethyl cellulose and polyvinyl alcohol. Dyepoly (ethylene glycol) has also been used for selective solubilisation of enzymes at such concentration of poly(ethylene glycol) which causes precipitation of proteins.
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References
Albertsson, P.- Å., Partition of Cell Particles and Macro molecules, Wiley, New York, 1986, 3rd edition.
Walter, H., Brooks, D.E. and Fisher, D. E. (eds.), Partitioning in Aqueous Two-Phase Systems. Theory, Methods, Uses, and Applications to Biotechnology, Academic Press, Orlando, 1985.
Johansson, G. and Joelsson, M., Liquid-liquid extraction of lactate dehydrogenase from muscle using polymer-bound triazine dyes. Appl. Biochem. Biotechnol., 1986, 13, 15–27.
Tjerneld, P., Johansson, G. and Joelsson, M., Affinity liquid-liquid extraction of lactate dehydrogenase on a large scale. Biotechnol. Bioeng. 1987, 30, 809–16.
Kroner, H.K., Cordes, A., Schelper, A., Morr, M., Buckmann, A.F. and Kula, M.-R., Affinity partition studied with glu-cose-6-phosphate dehydrogenase in aqueous two-phase systems in response to triazine dyes. In Chromatography and Related Techniques, eds., T.C.J. Gribnau, J. Visser and R.J.F. Nivard, Elsevier, Amsterdam, 1982, pp. 491–501.
Johansson, G., Kopperschläger, G. and Albertsson, P.-Å., Affinity partitioning of phosphofructokinase from baker’s yeast using polymer-bound Cibacron blue F3-A. Eur. J. Biochem., 1983, 131, 89–94.
Johansson, G. and Andersson, M., Parameters determining affinity partitioning of yeast enzymes using polymer-bound triazine dye ligands. J. Chromatogr. 1984, 303, 39–51.
Johansson, G., Aqueous two-phase systems in protein purification. J. Biotechnol., 1985, 3, 11–8.
Kopperschläger, G., Lorenz, G. and Usbeck, E., Application of affinity partitioning in an aqueous two-phase system to the investigation of triazine dye-enzyme interactions. J. Chromatogr., 1983, 259, 97–105.
Joelsson, M. and Johansson, G., Sequential liquid-liquid extraction of some enzymes from porcine muscle using polymer-bound triazine dyes, Enzyme Microb. Technol., 1987, 9, 233–7.
Bradford, M.M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of dye binding. Anal. Biochem., 1976, 72, 248–54.
Scopes, R.K., 3-Phosphoglycerate kinase of skeletal muscle. Methods Enzymol. 1975, 42, 127–34.
Noltmann, E.A., Gubler, C.J. and Kuby, S.A., Glucose 6-phosphate dehydrogenase (Zwischenferment). I. Isolation of crystalline enzyme from yeast. J. Biol. Chem., 1961, 236, 1225–30.
Bergmeyer, H.U., Methoden der enzymatischen Analyse, Verlag Chemie, Weinheim/Bergstr., 1970, 2nd ed., Vol. 1, p. 441.
Johansson, G. and Joelsson, M., Affinity partitioning of enzymes using dextran-bound Procion yellow HE-3G. Influence of dye-ligand density. J. Chromatogr., 1987, 393, 195–208.
Johansson, G. and Joelsson, M., Effect of polymer structure on affinity partitioning of lactate dehydrogenase in polymer-water two-phase systems. J. Chroinatogr., 1987, 411, 161–6.
Flanagan, S.D. and Barondes, S.H., Affinity partitioning — a method for purification of proteins using specific polymer-ligands in aqueous polymer two-phase systems. J. Biol. Chem., 1975, 250, 1484–9.
Cordes, A., Flossdorf, J. and Kula, M.-R., Affinity partitioning: development of mathematical model describing behaviour of biomolecules in aqueous two-phase systems. Biotechnol. Bioeng. 1987, 30, 514–20.
Johansson, G. and Joelsson, M., Specifically increased solubility of enzymes in polyethyleneglycol solutions using polymer-bound triazine dyes. Anal. Biochem., 1986, 158, 104–10.
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Johansson, G. (1989). Affinity Partition and Solubilisation of Enzymes by Use of Polymer-Bound Triazine Dyes. In: Vijayalakshmi, M.A., Bertrand, O. (eds) Protein-Dye Interactions: Developments and Applications. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-1107-9_18
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DOI: https://doi.org/10.1007/978-94-009-1107-9_18
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-010-6989-2
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