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Title 

Process development for production of recombinant human insulin-like growth factor-I in Escherichia coli

Authors 

Bong Hyun ChungY J ChoiS H YoonS Y LeeYoung Ik Lee

Publisher 

Springer

Issue Date 

2000

Citation 

Journal of Industrial Microbiology (& Biotechnology), vol. 24, no. 0, pp. 94-99

Keywords 

insulin-like growth factor I (IGF-I)fed-batch culturedownstream processingEscherichia coli

Abstract 

Fed-batch cultures were carried out to overproduce human insulin-like growth factor I (IGF-I) in Escherichia coli. The effects of carbon sources (glucose or glycerol) and induction time on cell growth and IGF-I production were investigated in more detail. Glycerol was a better carbon source than glucose for IGF-I production in fed-batch culture. Induction at the mid- exponential phase with glycerol as a carbon source in the pH-stat fed-batch culture was optimal for IGF-I production. Under this condition, 2.8 g L-1 of fusion IGF-I was produced as inclusion bodies. We have also developed downstream processing for preparative scale purification of IGF-I from the fusion protein produced by the fed-batch culture using glycerol as a carbon source. After the fusion protein expressed was solubilized in 8 M urea and cleaved with hydroxylamine, the released IGF-I was purified by cation exchange chromatography, refolding and preparative scale reverse phase HPLC (rp-HPLC) to give recombinant IGF-I of >98% purity. The biological activities of the purified IGF-I were measured and found to be identical to those of commercial IGF-I.

ISSN 

0169-4146

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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