Back to Search
Start Over
Temperature effect on the functional expression of human cytochromes P450 2A6 and 2E1 in Escherichia coli.
- Source :
-
Archives of pharmacal research [Arch Pharm Res] 2005 Apr; Vol. 28 (4), pp. 433-7. - Publication Year :
- 2005
-
Abstract
- Human cytochromes P450 (CYP) 2A6 and 2E1 are of great interest because of their important roles in the oxidation of numerous drugs and carcinogens. Bacterial expression systems, especially Escherichia coli cells, have been widely used for the production of various CYP enzymes in order to obtain high yield of proteins. The expression methods usually employ longer culture time (30-72 h) at lower temperature (usually under 30 degrees C). Expression levels of CYPs 2A6 and 2E1 at 37 degrees C were compared to those at 280 degrees C, which is a usual temperature used in most bacterial expression systems for human CYP expression. Within 18 h the expression levels of CYPs 2A6 and 2E1 reached up to 360 and 560 nmol per liter culture at 37 degrees C, respectively, which are compatible with those of 36 h culture at 280 degrees C. The activities of CYPs expressed at 37 degrees C were also comparable to those expressed at 28 degrees C. The present over-expression system can be useful for rapid production of large amounts of active human CYPs 2A6 and 2E1 in E. coli.
- Subjects :
- Aryl Hydrocarbon Hydroxylases isolation & purification
Aryl Hydrocarbon Hydroxylases metabolism
Cloning, Molecular
Cytochrome P-450 CYP2A6
Cytochrome P-450 Enzyme System isolation & purification
Cytochrome P-450 Enzyme System metabolism
Cytochrome P450 Family 2
Escherichia coli enzymology
Humans
Mixed Function Oxygenases isolation & purification
Mixed Function Oxygenases metabolism
Recombinant Proteins isolation & purification
Recombinant Proteins metabolism
Temperature
Aryl Hydrocarbon Hydroxylases biosynthesis
Cytochrome P-450 Enzyme System biosynthesis
Escherichia coli genetics
Mixed Function Oxygenases biosynthesis
Recombinant Proteins biosynthesis
Subjects
Details
- Language :
- English
- ISSN :
- 0253-6269
- Volume :
- 28
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Archives of pharmacal research
- Publication Type :
- Academic Journal
- Accession number :
- 15918516
- Full Text :
- https://doi.org/10.1007/BF02977672