Solubilization of the overexpressed integral membrane protein alkane monooxygenase of the recombinant Escherichia coli W3110[pGEc47].
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.
Institut für Biotechnologie, ETH-Hönggerberg, Zürich, Switzerland.
J
J
Peters
B
B
Witholt
D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry*; D042926:Cytochrome P-450 CYP4A; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics; D003902:Detergents; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics; D006899:Mixed Function Oxygenases / Q000737:chemistry* / Q000235:genetics; D011549:Pseudomonas / Q000235:genetics; D011995:Recombination, Genetic; D012995:Solubility
D001425:Bacterial Outer Membrane Proteins; D003902:Detergents; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D042926:Cytochrome P-450 CYP4A