Fermentation process optimization of recombinant Saccharomyces cerevisiae for the production of human interferon-alpha2a.
The effects of different culture conditions on the expression level of human interferon-alpha2a (IFN-alpha2a) by using recombinant yeast were investigated in a 2.6-L jar fermentor. Appropriate supplement of glucose and the maintenance of residual glucose at a low level resulted in the reduction of ethanol formation and enhancement of the bioactivity of IFN-alpha2a to 4.9 x 106 from 3.1 x 10(6) IU/mL. When adenine was added evenly for 10-20 h of fermentation into the basal culture medium at a speed of 2 microg/mL of medium/h, OD600 was greatly increased to 24, and the protein increased to 276 mg/L. The content of ethanol generated was also reduced tremendously during the process, and as a result, 1.3 x 10(7) IU/mL of biologic activity was achieved. In the expression phase, pH had an important impact on expression level, which should be controlled at 5.5.
Chu
Ju
J
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, PR China. juchu@online.sh.cn
Zhang
Siliang
S
Zhuang
Yingping
Y
D000225:Adenine / Q000494:pharmacology; D003470:Culture Media; D000431:Ethanol / Q000032:analysis / Q000378:metabolism; D005285:Fermentation; D005786:Gene Expression Regulation / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000235:genetics; D010461:Peptones / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D013997:Time Factors
D003470:Culture Media; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D010461:Peptones; D011994:Recombinant Proteins; D000431:Ethanol; D005947:Glucose; D000225:Adenine