Temperature effect on the functional expression of human cytochromes P450 2A6 and 2E1 in Escherichia coli.
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.
Yim
Sung-Kun
SK
School of Biological Sciences and Technology, Chonnam National University, Gwangju 500-757, Korea.
Ahn
Taeho
T
Jung
Heung-Chae
HC
Pan
Jae-Gu
JG
Yun
Chul-Ho
CH
D001189:Aryl Hydrocarbon Hydroxylases / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D065637:Cytochrome P-450 CYP2A6; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D000072467:Cytochrome P450 Family 2; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D006801:Humans; D006899:Mixed Function Oxygenases / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D013696:Temperature
D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D001189:Aryl Hydrocarbon Hydroxylases; C110332:CYP2F1 protein, human; D065637:Cytochrome P-450 CYP2A6; D000072467:Cytochrome P450 Family 2