Improving protein extraction yield in reversed micellar systems through surface charge engineering.
The mechanism of extraction of rat cytochrome b(5) from water into a sodium dioctylsulfosuccinate (AOT) micellar organic phase was studied using protein engineering of surface charged residues. The extraction behavior of native cytochrome b(5) and modified proteins with substitutions of the type glutamic acid --> lysine at positions 44 (E44K), 56 (E56K), and 92 (E92K), was studied as a function of pH. The results indicate that an important mechanism of extraction is an electrostatic interaction of this protein with the negatively charged surfactant. We demonstrate that it is possible to improve extraction by engineering the protein surface charge, increasing the driving force responsible for the protein transfer to the micellar phase. (c) 1994 John Wiley & Sons, Inc.
Pires
M J
MJ
Laboratório de Engenharia Bioquímica, Instituto Superior Técnico, 1000 Lisboa, Portugal.
Martel
P
P
Baptista
A
A
Petersen
S B
SB
Willson
R C
RC
Cabral
J M
JM