Impact of dissolved oxygen concentration on some key parameters and production of rhG-CSF in batch fermentation.
The impact of different levels of agitation speed, carbondioxide and dissolved oxygen concentration on the key parameters and production of rhG-CSF in Escherichia coli BL21(DE3)PLysS were studied. Lower carbondioxide concentrations as well as higher agitation speeds and dissolved oxygen concentrations led to reduction in the acetate concentrations, and enhanced the cell growth, but inhibited plasmid stability and rhG-CSF expression. Similarly, higher carbondioxide concentrations and lower agitation speeds as well as dissolved oxygen concentrations led to enhanced acetate concentrations, but inhibited the cell growth and protein expression. To address the bottlenecks, a two-stage agitation control strategy (strategy-1) and two-stage dissolved oxygen control strategy (strategy-2) were employed to establish the physiological and metabolic conditions, so as to improve the expression of rhG-CSF. By adopting strategy-1 the yields were improved 1.4-fold over constant speed of 550 rpm, 1.1-fold over constant dissolved oxygen of 45%, respectively. Similarly, using strategy-2 the yields were improved 1.6-fold over constant speed of 550 rpm, 1.3-fold over constant dissolved oxygen of 45%, respectively.
Krishna Rao
Dasari V
DV
Research and Development, Biotechnology Division, Natco Research Centre, Hyderabad, 500018, India. nrc@natcopharma.co.in
Ramu
Chatadi T
CT
Rao
Joginapally V
JV
Narasu
Mangamoori L
ML
Bhujanga Rao
Adibhatla Kali S
AK
D000085:Acetates / Q000378:metabolism; D001431:Bacteriological Techniques; D019149:Bioreactors; D002245:Carbon Dioxide / Q000494:pharmacology; D004926:Escherichia coli / Q000254:growth & development* / Q000378:metabolism*; D005285:Fermentation / Q000187:drug effects; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects*; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics*; D006801:Humans; D007218:Industrial Microbiology; D010100:Oxygen / Q000378:metabolism / Q000494:pharmacology*; D011994:Recombinant Proteins; D013995:Time; D014783:Viscosity
D000085:Acetates; D011994:Recombinant Proteins; D002245:Carbon Dioxide; D016179:Granulocyte Colony-Stimulating Factor; D010100:Oxygen