Intein-mediated expression and purification of an analog of glucagon-like peptide-1 in Escherichia coli.
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.
Ma
Chen
C
School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, P. R. China.
Gao
Mingming
M
Liu
Wenchao
W
Zhu
Jing
J
Tian
Hong
H
Gao
Xiangdong
X
Yao
Wenbing
W
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
D052216:Glucagon-Like Peptide 1