Abstract
As a promising type 2 anti-diabetic agent, glucagon-like peptide-1 (GLP-1) is attracting more and more interest. Mutated GLP-1 (mGLP-1) is an analog of native GLP-1. To facilitate the production and purification of mGLP-1, auto-induction and on-column cleavage was employed in this study. By using auto-induction system, after 24 h of shaking culture, about 12.6 g wet bacterial cells could be obtained from 1 l medium, and this was about 3.6 times more than that of the IPTG-induction group. After disruption and centrifugation, the fusion protein was directly purified and cleaved on Ni–Sepharose 6 Fast Flow column. Then, RESOURCE15 RPC column was used for further purification. By using these two steps of purification, about 1.58 mg of mGLP-1 with the purity of up to 98% could be obtained from 1 g wet bacterial cells. In the bioactivity study, mGLP-1 displayed a significant and dose-dependent glucose-lowering activity. These results suggested that auto-induction and on-column cleavage could facilitate the production and purification of mGLP-1. These methods could also be applied to the preparation of other proteins and peptides.
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Acknowledgments
This work was supported by the China National Nature Science Foundation (30772679, 30973667), the Hi-Tech Research and Development Program of China-863 Program (2007AA02Z101), and the innovation fund project of Simcere Pharmaceutical Group for graduate student at China Pharmaceutical University.
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Gao, M., Ma, C., Liu, W. et al. Production and purification of an analog of glucagon-like peptide-1 by auto-induction and on-column cleavage in Escherichia coli . World J Microbiol Biotechnol 26, 1675–1682 (2010). https://doi.org/10.1007/s11274-010-0345-3
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DOI: https://doi.org/10.1007/s11274-010-0345-3