Solubilization of the overexpressed integral membrane protein alkane monooxygenase of the recombinant Escherichia coli W3110[pGEc47]

Biochim Biophys Acta. 1994 Dec 30;1196(2):145-53. doi: 10.1016/0005-2736(94)00216-9.

Abstract

The integral membrane-bound alkane monooxygenase (AlkB) from Pseudomonas oleovorans has been overexpressed in the recombinant Escherichia coli strain W3110[pGEc47] and expression levels of 10 to 15% relative to the total cell protein were reached. The amount of phospholipids in induced cells is about 3-fold higher compared to the wild-type and AlkB has been shown to be located in small membrane vesicles. We present here a study on the solubilization of these AlkB containing membrane vesicles by different detergents with special emphasis on structural requirements for a surfactant preserving the activity of AlkB. Moreover, the effects of the detergents used on the complete alkane hydroxylase system was studied.

MeSH terms

  • Bacterial Outer Membrane Proteins / chemistry*
  • Cytochrome P-450 CYP4A
  • Cytochrome P-450 Enzyme System / chemistry*
  • Cytochrome P-450 Enzyme System / genetics
  • Detergents
  • Escherichia coli / chemistry*
  • Escherichia coli / genetics
  • Mixed Function Oxygenases / chemistry*
  • Mixed Function Oxygenases / genetics
  • Pseudomonas / genetics
  • Recombination, Genetic
  • Solubility

Substances

  • Bacterial Outer Membrane Proteins
  • Detergents
  • Cytochrome P-450 Enzyme System
  • Mixed Function Oxygenases
  • Cytochrome P-450 CYP4A