High-level expression of human cytochrome P450 1A2 by co-expression with human molecular chaperone HDJ-1(Hsp40).
Human cytochrome P450 (CYP) 1A2 is of great interest because of its important roles in the oxidation of numerous drugs and carcinogens. HDJ-1, a molecular chaperone in human, is known to assist the correct folding of unfolded proteins. To achieve high yield of recombinant human CYP1A2 in Escherichia coli, the CYP1A2 encoding gene was co-expressed with the chaperone HDJ-1 under the control of an inducible tac promoter in bicistronic format. Expression level of CYP1A2 in the bicistronic construct reached up to 520 nmol/liter culture within 16 h at 37 degrees C, which is 3.4-fold increase compared to the expression yield of CYP1A2 alone without HDJ-1. By co-expression with HDJ-1, the catalytic activity of CYP1A2 was also increased 5.5-fold. The activity increase seems to be associated with the increase of CYP production at whole cell level. The present over-expression system may be useful for rapid production of large amounts of active human CYP1A2 in E. coli.
Ahn
Taeho
T
Department of Biochemistry, College of Veterinary Medicine, Chonnam National University, Gwangju 500-757, Republic of Korea. thahn@chonnam.ac.kr
Yang
Siyoung
S
Yun
Chul-Ho
CH
D003001:Cloning, Molecular; D019388:Cytochrome P-450 CYP1A2 / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D050956:HSP40 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
C494647:DNAJB1 protein, human; D050956:HSP40 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D019388:Cytochrome P-450 CYP1A2