Lian, Ai Ai
(2019)
Salt-Induced Phase Separation Of
Chaps And Potential Application For
Hydrophobic Molecule Extraction.
Masters thesis, Universiti Sains Malaysia.
Abstract
CHAPS is a zwitterionic detergent commonly utilized in protein biochemistry
based applications. Despite widespread utility, understanding of CHAPS phase
behaviour remained poor. In this study, the effect of salts on CHAPS phase behaviour
was first investigated. Consolute solution temperature of CHAPS-(NH4)2SO4 systems
were determined, followed by phase diagram construction. Potential of CHAPS-(NH4-
)2SO4 systems for hydrophobic molecule extraction via phase separation was then
evaluated using Coomassie Brilliant Blue (CBB) and recombinant cytochrome P450
(CYP) 2C19. Sulphate and phosphate salts were successful in inducing phase
separation of CHAPS. CHAPS and (NH4)2SO4 systems exhibited an upper consolute
solution temperature (UCST). CBB was found to partition extensively to micelle-rich
phase (MRP) of the 4.5% CHAPS and 8% (NH4)2SO4 system with an extraction yield
of 83±0.32%. Extraction yield of recombinant CYP 2C19 in MRP of the 13.5%
CHAPS and 10.8% (NH4)2SO4 system was 46±7.58%. The outcomes of this study
propound the scope for application of CHAPS as a component of phase separation
systems.
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