Solubilization of enzymes in water-in-oil microemulsions and their rapid and efficient release through use of a pH-degradable surfactant |
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| Authors: | Maithili E Rairkar Douglas G Hayes J Milton Harris |
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| Institution: | (1) Department of Chemistry, University of Alabama in Huntsville, Huntsville, AL 35899, USA;(2) Present address: Discovery Laboratories, Inc., 3 350 N Bernardo Ave, Mountain View, CA 94043, USA;(3) Department of Chemical and Materials Engineering, University of Alabama in Huntsville, Huntsville, AL 35899, USA;(4) Nektar, Inc., 490 Discovery Drive, Huntsville, AL 35806, USA;(5) Department of Biosystems Engineering, University of Tennessee, 2506 E.J. Chapman Drive, Knoxville, TN 37996-4531, USA |
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| Abstract: | α-Chymotrypsin and lysozyme were solubilized in a water/O-(2-tridecyl, 2-ethyl-1,3-dioxolan-4-yl)methoxy]–O′-methoxy poly(ethylene glycol) (CK-2,13 surfactant)/isooctane water-in-oil microemulsion solution at 1.5–2 and 10 g l−1 for 0.15 and 1.2 M CK-2,13, respectively. Upon contact with an equal volume of 0.1 M NaH2PO4/Na2HPO4 buffer, pH 5, a three-phase system (Winsor-III system) was formed, consisting of a surfactant-rich middle phase and aqueous
and isooctane-rich “excess” phases. Both enzymes were rapidly released into the aqueous excess phase, with 70% recovery of
each in 30 and 60 min for microemulsion solutions containing 0.15 and 1.2 M surfactant, respectively. The recovered enzymes retained >90% of their original specific activity. |
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| Keywords: | pH-degradable surfactant Protein extraction (purification) Protein solubilization Reverse micelles Water-in-oil microemulsions |
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