[Hepatitis E virus capsid protein production by high cell density culture of recombinant Escherichia coli].
Production of Hepatitis E Virus capsid protein by high cell density culture in recombinant E. coli has been studied in 10L and 30L fermentors. The effects of different factors on growth and producing recombinant protein of E. coli have been studied by batch culture, such as different media, the ratio of phosphate and Magnesium sulfate. Comparison of fermentation performance for recombinant E. coli in different fed-methods culture has been investigated by fed-batch culture. The effects of inducing at different stages of growth and time of inducing on growth and producing recombinant protein, also obtained by fed-batch culture. At last, the solubility of inclusion body in different urea concentrations also has been obtained by fed-batch culture. The results show that the concentration of phosphate and Magnesium sulfate in the optimal media is 80mmol/L and 20mmol/L in batch culture respectively, that induction with 1.0mmol/L IPTG at mid log phase (about 45 OD at 600nm) is suitable for growth and recombinant protein expression, the cells were approaching stationary growth phase and the maximum cell OD at 600nm of 80 was achieved in 5h of fed-batch culture, and the expression level is 29.74%. The results also indicate that the solubility of inclusion body in 4mol/L urea solution induced at 37 degrees C reaches 14mg/mL, over 80% inclusion body was resolved. The culture process achieved in 10L fermentor could be successfully scaled up to 30L fenmentor with good reproducibility.
Liu
Ru-Shi
RS
The Key Laboratory of the Ministry of Education for Cell Biology and Tumor Cell Engineering, Xiamen 361005, China.
He
Zhi-Qiang
ZQ
Li
Shao-Wei
SW
Yang
Kun-Yu
KY
Xian
Yang-Ling
YL
Pang
Shu-Qiang
SQ
Zhang
Jun
J
Li
Yi-Min
YM
Xia
Ning-Shao
NS
D019149:Bioreactors / Q000382:microbiology; D015169:Colony Count, Microbial; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D016752:Hepatitis E virus / Q000235:genetics*; D019590:Nucleocapsid Proteins / Q000096:biosynthesis* / Q000235:genetics; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics
D019590:Nucleocapsid Proteins; D011993:Recombinant Fusion Proteins; C099600:nucleocapsid protein, Hepatitis virus