Solubilization and activity of proteins in compressible-fluid based microemulsions.
We have used the pressure dependency of density in near critical propane to design a novel protein extraction system. Active, structurally intact, proteins have been extracted from an aqueous phase into subcritical propane containing the non-ionic detergent polyoxyethylene sorbitan trioleate (Tween-85). The pressure dependence of protein transfer from the aqueous to organic phase is described for cytochrome C and subtilisin. The effect of the microemulsion and compressible propane on protein structure and function is also discussed. The first determinations of kinetic constants for an enzyme-catalyzed reaction in such a system are also presented.
Ayala
G
G
Department of Chemical Engineering, University of Pittsburgh, Pennsylvania 15261.
Kamat
S V
SV
Komives
C
C
Beckman
E J
EJ
Russell
A J
AJ
D003574:Cytochrome c Group / Q000737:chemistry / Q000302:isolation & purification; D004655:Emulsions; D011136:Polysorbates; D011312:Pressure; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012995:Solubility; D012996:Solutions; D013381:Subtilisins / Q000737:chemistry / Q000302:isolation & purification
D003574:Cytochrome c Group; D004655:Emulsions; D011136:Polysorbates; D011506:Proteins; D012996:Solutions; D013381:Subtilisins