Production of a platelet aggregation inhibitor, salmosin, by high cell density fermentation of recombinant Escherichia coli.
Optimal conditions for a high cell density fermentation were investigated in a recombinant Escherichia coli producing salmosin, a platelet aggregation inhibitor. The optimized carbon and nitrogen sources were glycerol 10 g/l, yeast extract 30 g/l, and bacto-tryptone 10 g/l, yielding the dry cell weight (DCW) of 10.61 g/l in a 500 ml flask culture. The late-stage induction with 1% L-arabinose in a 5 l jar fermentor showed the highest DCW of 65.70 g/l after 27 h of the fed-batch fermentation. Around 2,200 mg/l of the protein was expressed as an inclusion body that was then refolded to obtain the active salmosin of 96 mg/l. We also confirmed the inhibitory activity against platelet aggregation of the active salmosin from the high cell density fermentation.
Department of Chemistry, Box H, Brown University, Providence, RI 02912-9108, USA.
Myung-Ji
MJ
Seo
Hak-Jong
HJ
Choi
Kwang-Hoe
KH
Chung
Yu-Ryang
YR
Pyun
D061249:Batch Cell Culture Techniques / Q000379:methods*; D003435:Crotalid Venoms / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D005285:Fermentation; D015870:Gene Expression; D006801:Humans; D010974:Platelet Aggregation / Q000187:drug effects; D010975:Platelet Aggregation Inhibitors / Q000378:metabolism* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
D003435:Crotalid Venoms; D010975:Platelet Aggregation Inhibitors; D011994:Recombinant Proteins; C115393:salmosin