High-level expression of fusion protein containing 10 tandem repeated GLP-1 analogs in yeast Pichia pastoris and its biological activity in a diabetic rat model.
Glucagon-like peptide-1 (GLP-1), an incretin secreted by intestinal L-cells, can effectively lower blood glucose levels in patients with diabetes. A fusion gene, consisting of 10 tandem repeated GLP-1 analog genes, was expressed at a high level in the yeast Pichia pastoris. SDS polyacrylamide gel electrophoresis (SDS-PAGE), and Western Blotting results showed that fusion protein migrated as a single protein band with a molecular weight of 36 kDa. A biological activity test showed that the GLP-1 analog could significantly lower the level of serum glucose when GLP-1 purified analog was injected into diabetic rats. A potential strategy for large-scale production of fusion protein containing the 10 GLP-1 analogs as discovered, and a single GLP-1 analog was obtained from fusion protein digested with trypsin. This should be inspired foreign expression of medicinal short peptides and be valuable in further research on GLP-1 analog drugs in the treatment of diabetes mellitus.
Hou
Jianhua
J
The Key Laboratory for Bioactive Materials of the Ministry of Education, Institute for Molecular Biology, College of Life Science, Nankai University, P. R. China.
Yan
Ruixiang
R
Yang
Liquan
L
Wu
Zhiqiang
Z
Wang
Cuiyan
C
Ding
Dongfeng
D
Li
Na
N
Ma
Chunxiao
C
Li
Minggang
M
D000818:Animals; D001786:Blood Glucose / Q000187:drug effects; D003001:Cloning, Molecular / Q000379:methods*; D003920:Diabetes Mellitus / Q000188:drug therapy*; D004195:Disease Models, Animal; D052216:Glucagon-Like Peptide 1 / Q000031:analogs & derivatives / Q000235:genetics* / Q000627:therapeutic use*; D010843:Pichia / Q000235:genetics*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000627:therapeutic use; D020080:Tandem Repeat Sequences
D001786:Blood Glucose; D011993:Recombinant Fusion Proteins; D052216:Glucagon-Like Peptide 1