Production of tilapia insulin-like growth factor-2 in high cell density cultures of recombinant Escherichia coli.

摘要: An improved expression plasmid pET-insulin-like growth factor-2 (IGF2) was constructed and transferred into Escherichia coli BL21(DE3) for the of tilapia insulin-like factor-2. The recombinant produced as inclusion bodies, content high 10.3% total protein content. For production in E. coli, pH-stat fed-batch cultures were used to achieve a cell density culture. A concentration 183gl(-1) dry weight (DCW) obtained after 30h cultivation stability maintained at levels. Expression induced three different concentrations, 50, 78.5, 114.5gl(-1) weight. When cells weight, amount 9.69gl(-1) (11.3% protein). Using simple purification process including body isolation, denaturation, refolding Ni-NTA affinity chromatography, 19.51mg from 22.5ml culture, recovery yield 20.5%. biological activity purified IGF-2 demonstrated promoting four lines by colorimetric bioassay best stimulation ratio Balb/3T3 clone 31A line.