Fermentation and purification of LacZ-fused single chain insulin precursor for (B3-homoserine) human insulin

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Title
Fermentation and purification of LacZ-fused single chain insulin precursor for (B3-homoserine) human insulin
Author(s)
S Y Lee; Jeong Heon Ko; M H Choi; D H Nam
Bibliographic Citation
Biotechnology and Bioprocess Engineering, vol. 1, no. 1, pp. 9-12
Publication Year
1996
Abstract
In order to produce the single chain precursor of a novel human insulin analogue, (B30-Homoserine) insulin, the fermentative behaviors of Escherichia coli JM103 were studied, which harbors pKBA plasmid carrying a hybrid gene in which the gene for a single chain precursor was fused with lacZ gene under tac promoter. The maximal induction of gene expression was achieved when more than 0.05 mM of isopropyl- β -D-thiogalactopyranoside(IPTG) was supplemented to fermentation medium after 4 h cultivation of E. coli, and followed by longer than 2-h fermentation. The hybrid protein of the single chain insulin precursor was isolated from cytoplasmic inclusion bodies by dissolving in 8M urea solution, and purified through DEAE-Sephacel and Sephadex G-200 column chromatographies with a recovery of 35%. The finally purified hybrid protein showed a single band on sodium dodecyl sulfate-polyacrylamide gel.
Keyword
Recombinant insulinInsulin analogue(B30-homoserine) insulinSingle chain insulin precursorGene inductiontac promoterInclusion body
ISSN
1226-8372
Publisher
Springer
Type
Article
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Genome Editing Research Center > 1. Journal Articles
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