Improving the yield of soluble 6xHis-tagged interferon-alpha via the addition of repressor of the araBAD promoter system in Escherichia coli.
The inhibition of inclusion body formation in Escherichia coli by the addition of alpha-D-glucopyranoside or D-fucose after induction improved the purification yield of soluble recombinant interferon-alpha. When D-fucose was added after induction, more soluble 6xHis-tagged interferon-alpha could be purified compared to when methyl alpha-D-glucopyranoside was added. It was shown that, on the basis of 1 mg dry cell weight, 16.6 microg of soluble 6xHis-tagged interferon-alpha was purified when D-fucose was added after induction and 6 ml nickel-chelated agarose gel column was used. This was about 15 times greater than when induction only was performed and 1 ml nickel-chelated agarose gel was used.
Jung
Kyung-Hwan
KH
Division of Food and Biotechnology, Chungju National University, Jeungpyung, Chungbuk 368-701, Republic of Korea. khjung@cjnu.ac.kr
Park
Young-Seo
YS
Yeon
Ji-Hyeon
JH
Kim
Sung-Hun
SH
Yoo
Sun Kyun
SK
Jang
Byeong-Churl
BC
D015153:Blotting, Western; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005990:Glycerol / Q000378:metabolism; D006639:Histidine / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism; D016898:Interferon-alpha / Q000302:isolation & purification / Q000378:metabolism*; D009842:Oligopeptides / Q000302:isolation & purification / Q000378:metabolism*; D009876:Operon / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism*; D012995:Solubility
C471213:His-His-His-His-His-His; D016898:Interferon-alpha; D009842:Oligopeptides; D011994:Recombinant Proteins; D012097:Repressor Proteins; D006639:Histidine; D005990:Glycerol