Abstract
To facilitate expression and purification of an analog of GLP-1 (mGLP-1), an intein system was employed in this study. A recombinant fusion protein, CBD-DnaB-mGLP-1, was constructed and expressed in the form of inclusion body. After refolding, the intein-mediated self-cleavage was triggered by pH and temperature shift. By using chitin beads column followed by single step purification, about 2.58 mg of mGLP-1 with the purity of up to 98% could be obtained from 1 L medium. Tricine-SDS-PAGE, RP-HPLC, and ESI-MS were undertaken to determine the purity and molecular weight of mGLP-1. The glucose-lowering activity of mGLP-1 was also preliminarily determined.
Keywords: Chitin beads, expression and purification, glucagon-like peptide-1, incretin, intein, self-cleavage
Protein & Peptide Letters
Title: Intein-Mediated Expression and Purification of an Analog of Glucagon-like Peptide-1 in Escherichia coli
Volume: 17 Issue: 10
Author(s): Chen Ma, Mingming Gao, Wenchao Liu, Jing Zhu, Hong Tian, Xiangdong Gao and Wenbing Yao
Affiliation:
Keywords: Chitin beads, expression and purification, glucagon-like peptide-1, incretin, intein, self-cleavage
Abstract: To facilitate expression and purification of an analog of GLP-1 (mGLP-1), an intein system was employed in this study. A recombinant fusion protein, CBD-DnaB-mGLP-1, was constructed and expressed in the form of inclusion body. After refolding, the intein-mediated self-cleavage was triggered by pH and temperature shift. By using chitin beads column followed by single step purification, about 2.58 mg of mGLP-1 with the purity of up to 98% could be obtained from 1 L medium. Tricine-SDS-PAGE, RP-HPLC, and ESI-MS were undertaken to determine the purity and molecular weight of mGLP-1. The glucose-lowering activity of mGLP-1 was also preliminarily determined.
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Cite this article as:
Ma Chen, Gao Mingming, Liu Wenchao, Zhu Jing, Tian Hong, Gao Xiangdong and Yao Wenbing, Intein-Mediated Expression and Purification of an Analog of Glucagon-like Peptide-1 in Escherichia coli, Protein & Peptide Letters 2010; 17 (10) . https://dx.doi.org/10.2174/092986610792231582
DOI https://dx.doi.org/10.2174/092986610792231582 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
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