Enhanced production of recombinant streptokinase in Escherichia coli using fed-batch culture.
Fed-batch culture strategy is often used for increasing production of heterologous recombinant proteins in Escherichia coli. This study was initiated to investigate the effects of dissolved oxygen concentration (DOC), complex nitrogen sources and pH control agents on cell growth and intracellular expression of streptokinase (SK) in recombinant E. coli BL21(DE3). Increase in DOC set point from 30% to 50% did not affect SK expression in batch culture where as similar increase in fed-batch cultivation led to a significant improvement in SK expression (from 188 to 720 mg l(-1)). This increase in SK could be correlated with increase in plasmid segregational stability. Supplementation of production medium with yeast extract and tryptone and replacement of liquid ammonia with NaOH as pH control agent further enhanced SK expression without affecting cell growth. Overall, SK concentration of 1120 mg l(-1) representing 14-fold increase in SK production on process scale-up from flask to bioreactor scale fed-batch culture is the highest reported concentration of SK to date.
Goyal
Deepika
D
Institute of Microbial Technology, Council of Scientific and Industrial Research, Chandigarh, India.
Sahni
Girish
G
Sahoo
Debendra K
DK
D000667:Ampicillin / Q000494:pharmacology; D019149:Bioreactors / Q000382:microbiology*; D003470:Culture Media; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D009584:Nitrogen / Q000494:pharmacology; D010100:Oxygen / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012995:Solubility / Q000187:drug effects; D013300:Streptokinase / Q000096:biosynthesis*
D003470:Culture Media; D011994:Recombinant Proteins; D000667:Ampicillin; D013300:Streptokinase; D009584:Nitrogen; D010100:Oxygen