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

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Title
Process development for production of recombinant human insulin-like growth factor-I in Escherichia coli
Author(s)
Bong Hyun Chung; Y J Choi; Sung Ho Yoon; S Y Lee; Young Ik Lee
Bibliographic Citation
Journal of Industrial Microbiology & Biotechnology, vol. 24, pp. 94-99
Publication Year
2000
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.
Keyword
insulin-like growth factor I (IGF-I)fed-batch culturedownstream processingEscherichia coli
ISSN
0169-4146
Publisher
Springer
DOI
http://dx.doi.org/10.1038/sj.jim.2900773
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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