Stable colloidal dispersions of a lipase-perfluoropolyether complex in liquid and supercritical carbon dioxide.
The technique of hydrophobic ion pairing was used to solubilize the lipase from Candida rugosa in a fluorinated solvent, perfluoromethylcyclohexane (PFMC), in complex with a perfluoropolyether (PFPE) surfactant, KDP 4606. The enzyme-surfactant complex was determined to have a hydrodynamic diameter of 6.5 nm at atmospheric pressure by dynamic light scattering (DLS), indicating that a single lipase molecule is stabilized by surrounding surfactant molecules. The complex formed a highly stable colloidal dispersion in both liquid and supercritical carbon dioxide at high CO2 densities (>0.92 and 0.847 g/mL, respectively), with 4% by volume PFMC as a cosolvent, yielding a fluid that was orange, optically translucent, and very nearly transparent. DLS demonstrated aggregation of the enzyme-surfactant complexes in CO2 at 25 and 40 degrees C and various pressures (2000-5000 psia) with hydrodynamic diameters ranging from 50 to 200 nm. The mechanism by which the enzyme-surfactant particles aggregate was shown to be via condensation due to very low polydispersities as characterized by the size distribution moments. Interparticle interactions were investigated with respect to density and temperature, and it was shown that on decreasing the CO2 density, the particle size increased, and the stability against settling decreased. Particle size also decreased as the temperature was increased to 40 degrees C, at constant CO2 density. Nanoparticle aggregates of an enzyme-surfactant complex in CO2, which are nearly optically transparent and stable to settling, are a promising new alternative to previous types of dispersions of proteins in CO2 that either required water/CO2 microemulsions or were composed of large particles unstable to settling.
Adkins
Stephanie S
SS
Department of Chemical Engineering, The University of Texas at Austin, Austin, Texas 78712, USA.
Hobbs
Helen R
HR
Benaissi
Karima
K
Johnston
Keith P
KP
Poliakoff
Martyn
M
Thomas
Neil R
NR
D002175:Candida / Q000201:enzymology; D002245:Carbon Dioxide / Q000737:chemistry*; D003102:Colloids / Q000737:chemistry*; D004987:Ethers / Q000737:chemistry*; D005466:Fluorocarbons / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D008027:Light; D008049:Lipase / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry; D011312:Pressure; D012542:Scattering, Radiation; D012995:Solubility; D012997:Solvents / Q000737:chemistry; D013499:Surface Properties; D013501:Surface-Active Agents / Q000737:chemistry*; D013997:Time Factors
D003102:Colloids; D004987:Ethers; D005466:Fluorocarbons; D012997:Solvents; D013501:Surface-Active Agents; C078113:perfluoropolyether; D002245:Carbon Dioxide; D008049:Lipase; C003073:perfluoromethylcyclohexane