Studies on the extraction of DesPro(2)-Val15-Leu17-aprotinin from the culture broth of a recombinant Saccharomyces cerevisiae.
The application of an aqueous two-phase system is described for the extraction of DesPro(2)-Val15-Leu17-aprotinin from yeast culture supernatant, using native chymotrypsin as affinity ligand. The interaction is driven by hydrophobic forces and leads to the accumulation of the aprotinin-chymotrypsin complex in the salt-rich (bottom) phase of a polyethyleglycol/salt system. The complex may be dissociated at low pH values. The separation of the recombinant aprotinin and the protease required chromatographic processes, which proved difficult to interface with the affinity extraction.
Barthel
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Institut für Enzymtechnologie, Universität Düsseldorf, Jülich, Germany.
Kula
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MR
D007611:Aprotinin / Q000031:analogs & derivatives* / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D002918:Chymotrypsin; D003470:Culture Media; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D011092:Polyethylene Glycols; D011994:Recombinant Proteins / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae; D012492:Salts
D003470:Culture Media; D007202:Indicators and Reagents; D008024:Ligands; D011994:Recombinant Proteins; D012492:Salts; C085595:aprotinin, desprolyl(2)-valyl(15)-leucyl(17)-; D011092:Polyethylene Glycols; D007611:Aprotinin; D002918:Chymotrypsin