Purification and bioactivity of exendin-4, a peptide analogue of GLP-1, expressed in Pichia pastoris.
Exendin-4, a peptide analogue of glucagon-like peptide-1 (GLP-1), has been developed for treatment of type 2 diabetes. Herein, the secretive exendin-4 fusion protein, expressed by methanol induction in Pichia pastoris system, was purified to homogeneity by chromatography followed by enterokinase cleavage of the fusion protein and subsequent purification of the recombinant exendin-4. Purity of the recombinant exendin-4 was 95.6%. Bioactivity assay revealed that it had glucose-lowering and insulin-releasing action in vivo.
State Key Laboratory of Bioreactor Engineering, National Engineering Research Center for Biotechnology (Shanghai), East China University of Science & Technology, Shanghai 200237, P.R. China.
Jin
J
Zhou
Ju
J
Chu
Yong-Hong
YH
Wang
Hui
H
Wang
Ying-Ping
YP
Zhuang
Si-Liang
SL
Zhang
D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D000077270:Exenatide; D052216:Glucagon-Like Peptide 1 / Q000031:analogs & derivatives*; D015151:Immunoblotting; D007274:Injections, Intraperitoneal; D007328:Insulin / Q000097:blood; D008297:Male; D010455:Peptides / Q000008:administration & dosage / Q000235:genetics / Q000302:isolation & purification*; D010843:Pichia / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000302:isolation & purification* / Q000378:metabolism; D014688:Venoms / Q000008:administration & dosage / Q000235:genetics / Q000302:isolation & purification*
D001786:Blood Glucose; D007328:Insulin; D010455:Peptides; D011994:Recombinant Proteins; D014688:Venoms; D052216:Glucagon-Like Peptide 1; D000077270:Exenatide