Expression and characterization of Momordica Chanrantia anti-hyperglycaemic peptide in Escherichia coli.
A nucleic acid sequence MC, encoding Momordica Chanrantia anti-hyperglycaemic peptide MC6 (accession: AAX06814) synthesized according to Escherichia coli preferred codons, was cloned and expressed in E. coli. Recombinant protein pQE8-MC (about 3.5 kDa) was purified and analyzed by 20% SDS-PAGE and western blot. It revealed that the expressed pQE8-MC had good solubility in aqueous media. An HPLC assay was used to confirm the expression of pQE8-MC. Subsequent pharmacological activity assay revealed a significant hypoglycemic effect of low dose treatments of pQE8-MC on male kunming mice. Four hours after an intravenous tail injection, the blood sugar levels of mice treated with pQE8-MC saline solution A3 (1 mg/kg BW) decreased greatly (P < 0.01) relative to the levels of a control group. This suggests that pQE8-MC, expressed in bioengineered E. coli, has a similar hypoglycemic function to the natural protein MC6 from M. Chanrantia. These results reveal the possibility of using bio-engineered bacteria as an anti-diabetic agent.
Liu
Si-Xiu
SX
Department of Environmental Science & Engineering, Urban Eco-Planning & Design Researching Center, Fudan University, 200433, Shanghai, People's Republic of China. liusixiu@gmail.com
Fu
Zhong-Ping
ZP
Mu
Rui-Min
RM
Hu
Zhi-Bi
ZB
Wang
Fu-Jun
FJ
Wang
Xiang-Rong
XR
D000496:Alloxan; D000818:Animals; D001786:Blood Glucose / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000188:drug therapy; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D006943:Hyperglycemia / Q000188:drug therapy*; D008297:Male; D051379:Mice; D029775:Momordica / Q000737:chemistry*; D010455:Peptides / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology / Q000627:therapeutic use*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology / Q000627:therapeutic use
D001786:Blood Glucose; D010455:Peptides; D011994:Recombinant Proteins; D000496:Alloxan