High level production of bovine angiogenin in E. coli by an efficient refolding procedure.
Recombinant bovine angiogenin (rbAng) was expressed in E. coli at up to 30% of total cell proteins but was produced as inclusion bodies. By investigating the effect of various factors on the refolding yield, we obtained about 60% refolding. After chromatographic purification, about 60 mg purified angiogenin was obtained from 1 l culture. The purified recombinant bovine angiogenin was identical to native bovine angiogenin (nbAng) obtained from cow's milk. Our approach is highly efficient and can be generally used for the production of various types of angiogenin for functional and structural studies as well as therapeutic purposes.
Jang
Seung-Hwan
SH
Department of Bioinformatics and Life Science, and CAMDRC, Soongsil University, Seoul 156-743, Korea.
Kang
Dong-Ku
DK
Chang
Soo-Ik
SI
Scheraga
Harold A
HA
Shin
Hang-Cheol
HC
D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D002645:Chickens; D049033:Chorioallantoic Membrane / Q000098:blood supply / Q000187:drug effects; D042783:Endothelial Cells / Q000187:drug effects / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D015202:Protein Engineering / Q000379:methods*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D012259:Ribonuclease, Pancreatic / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D014945:Wound Healing / Q000187:drug effects
D011994:Recombinant Proteins; C046366:angiogenin; D012259:Ribonuclease, Pancreatic