Improving protein extraction yield in reversed micellar systems through surface charge engineering

M. J. Pires; P. Martel; A. Baptista; S. B. Petersen; R. C. Willson; J. M.S. Cabral

doi:10.1002/bit.260440702

Improving protein extraction yield in reversed micellar systems through surface charge engineering

M. J. Pires, P. Martel, A. Baptista, S. B. Petersen, R. C. Willson, J. M.S. Cabral

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

The mechanism of extraction of rat cytochrome b₅ 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 b₅ 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.

Original language	English (US)
Pages (from-to)	773-780
Number of pages	8
Journal	Biotechnology and Bioengineering
Volume	44
Issue number	7
DOIs	https://doi.org/10.1002/bit.260440702
State	Published - Sep 20 1994

Keywords

Liquid-liquid extraction
Protein engineering
Reverse micelles

ASJC Scopus subject areas

Biotechnology
Microbiology

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10.1002/bit.260440702

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title = "Improving protein extraction yield in reversed micellar systems through surface charge engineering",

abstract = "The mechanism of extraction of rat cytochrome b5 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 b5 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.",

keywords = "Liquid-liquid extraction, Protein engineering, Reverse micelles",

author = "Pires, {M. J.} and P. Martel and A. Baptista and Petersen, {S. B.} and Willson, {R. C.} and Cabral, {J. M.S.}",

year = "1994",

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T1 - Improving protein extraction yield in reversed micellar systems through surface charge engineering

AU - Pires, M. J.

AU - Martel, P.

AU - Baptista, A.

AU - Petersen, S. B.

AU - Willson, R. C.

AU - Cabral, J. M.S.

PY - 1994/9/20

Y1 - 1994/9/20

N2 - The mechanism of extraction of rat cytochrome b5 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 b5 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.

AB - The mechanism of extraction of rat cytochrome b5 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 b5 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.

KW - Liquid-liquid extraction

KW - Protein engineering

KW - Reverse micelles

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JO - Biotechnology and Bioengineering

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Improving protein extraction yield in reversed micellar systems through surface charge engineering

Abstract

Keywords

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