title: Intein-mediated expression and purification of an analog of glucagon-like peptide-1 in Escherichia coli.
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.
authors.affiliation: School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, P. R. China.
authors.forename: Chen
authors.initials: C
authors.lastname: Ma
authors.forename: Mingming
authors.initials: M
authors.lastname: Gao
authors.forename: Wenchao
authors.initials: W
authors.lastname: Liu
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.forename: Hong
authors.initials: H
authors.lastname: Tian
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Gao
authors.forename: Wenbing
authors.initials: W
authors.lastname: Yao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D052216:Glucagon-Like Peptide 1 / Q000031:analogs & derivatives* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D047668:Inteins; D051379:Mice; D008969:Molecular Sequence Data
chemical_list: D052216:Glucagon-Like Peptide 1