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

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

Research output: Contribution to journalArticle

12 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 languageEnglish (US)
Pages (from-to)773-780
Number of pages8
JournalBiotechnology and Bioengineering
Volume44
Issue number7
DOIs
StatePublished - Jan 1 1994

Keywords

  • Liquid-liquid extraction
  • Protein engineering
  • Reverse micelles

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

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