Abstract
Downstream processing of bioproducts results in considerable losses of compounds of interest in a large number of cases. For the intracellular enzyme tartrate dehydrogenase, an analysis of the laboratory process for enzyme recovery revealed that maximum losses occur in the initial stages of purification when the enzyme is separated from nucleic acids and other undesirable enzymes. Hence, aqueous twophase extraction was studied to investigate the separation of several enzymes from nucleic acids. Single-component and binary equilibria for three commercially available enzymes (bovine serum albumin, trypsin, chymotrypsin) and yeast RNA were studied in a two-phase system consisting of dextran and polyethylene glycol (PEG). The effects of pH and concentrations of the components and salts (NaC1) were investigated.
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References
Polson, A. (1953),Biochim. Biophys. Acta 11, 315.
Largier, J. F. (1956),Biochim. Biophys Acta 21, 433.
Largier, J. F. (1957),J. Immunol. 79, 181.
Mathies, J. C. (1952),Science 115, 144.
Oxenburgh, M. S. and Snoswell, A. N. (1965),Nature 203, 1416.
Treybal, R. E. (1963),Liquid Liquid Extraction. McGraw Hill, New York.
Albertsson, P.-Å. (1965),Nature (Lond.)177, 771.
Albertsson, P.-Å. (1985), inPartitioning in Aqueous Two Phase Systems: Theory, Methods, Uses and Applications in Biotechnology, Walter H., Brooks, D. E., and Fisher D., eds., Academic, New York, p. 1.
Johansson, G. and Joelsson, M. (1989), inSeparations Using Aqueous Two Phase Systems, Fisher, D. and Sutherland, I. A., eds., Plenum, New York, p. 33.
Mattiasson, B. (1983),Trends Biotechnol. 1, 16.
Kula, M.-R., Kroner, K. H., and Huesdt, H. (1982),Adv. Biochem. Eng. 24, 73.
Kroner, K. H., Cordes, A., Flossdorf, J., and Kula, M.-R. (1982), inAffinity Chromatography and Related Techniques, Gribnau T. C. J. and Nivard R. J. F., eds., Elsevier, Amsterdam, p. 491.
Johansson, G. (1974),Acta Chem. Scand. Ser. 28, 872.
Cascone, O., Andrews, B. A., and Asenjo, J. A. (1991),Enzyme Microb. Technol. 13, 829.
Tipton, P. A. and Peisach, J. (1990),Biochemistry 29, 1749.
Kopperschlager, G. and Johansson, G. (1982),Anal. Biochem. 124, 117.
Bamberger, S., Seaman, G. V. F., Sharp, K. A., and Brooks, D. E. (1984),J. Colloid Interface Sci. 99, 187.
Warburg, O. and Christian, W. (1941),Biochem. Z. 310.
Albertsson, P.-Å. (1971),Partition of Cell Particles and Macromolecules, Wiley, New York, p. 99.
Fasman, G. D. (1977),Handbook of Biochemistry and Molecular Biology, 3rd ed., CRC, Cleveland, OH.
Johansson, G. (1974),Mol. Cell. Biochem. 4 (3), 169.
Szlag, D. C. and Guiliano, K. A. (1988),Biotechnol. Techniques 2, 4, 277.
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Bajpai, R., Harve, R. & Tipton, P. Effect of nucleic acid contamination on partitioning of proteins in two-phase PEG-Dextran system. Appl Biochem Biotechnol 54, 193–205 (1995). https://doi.org/10.1007/BF02787919
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DOI: https://doi.org/10.1007/BF02787919