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Expression and characterization of Momordica Chanrantia anti-hyperglycaemic peptide in Escherichia coli

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Abstract

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.

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Acknowledgments

We acknowledge associate professor Fujun wang (Institute traditional Chinese Medicine, shanghai university of traditional Chinese medicine), and associate professor Juan Lin (State Key Laboratory of Genetic Engineering, Fudan University) for providing pMD18-MC, pQE8 and M15, respectively. This work was supported by a grant (no. 04DZ19818) from Shanghai Municipal Commission of Science and Technology Department.

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Authors and Affiliations

  1. Department of Environmental Science & Engineering, Urban Eco-Planning & Design Researching Center, Fudan University, 200433, Shanghai, People’s Republic of China

    Si-Xiu Liu, Rui-Min Mu & Xiang-Rong Wang

  2. Institute of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, 201203, Shanghai, China

    Si-Xiu Liu, Zhong-Ping Fu, Zhi-Bi Hu & Fu-Jun Wang

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  1. Si-Xiu Liu
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Correspondence to Si-Xiu Liu.

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Liu, SX., Fu, ZP., Mu, RM. et al. Expression and characterization of Momordica Chanrantia anti-hyperglycaemic peptide in Escherichia coli . Mol Biol Rep 37, 1781–1786 (2010). https://doi.org/10.1007/s11033-009-9609-0

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  • DOI: https://doi.org/10.1007/s11033-009-9609-0

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