A study on refolding recombinant human leptin protein from Escherichia coli inclusion body

Authors

  • Le Mai Huong Xuan University of Science, VNU-HCM
  • Dang Thi Phuong Thao University of Science, VNU-HCM
  • Tran Linh Thuoc University of Science, VNU-HCM

DOI:

https://doi.org/10.15625/0866-7160/v36n1se.4365

Keywords:

Escherichia coli, Agglutination, inclusion body, leptin, refolding, solulibization

Abstract

Leptin, a peptide hormone, is produced by mature adipocytes and functions primarily in the hypothalamus in regulating food intake and body metabolism. We have studied the production of recombinant human leptin in the bacterial system Escherichia coli for generating high-quality materials for the study on development of  body weight regulation products in human. It was found that high-level expression of recombinant leptin protein in E. coli led to the formation of inclusion bodies. Therefore, solubilization and refolding leptin in inclusion bodies are crucial for the production of recombinant leptin from E. coli. Our results showed that leptin inclusion bodies can be solubilized by low concentration of denaturant, 2 M urea solution pH 12.5. Then, the protein was refolded by using dilution method or dialysis method. The native form of proteins  has been confirmed by SDS-PAGE under reducing and non-reducing conditions, Native-PAGE, agglutination absortion at OD340; and the concentration was also determined by Bradford method. The recovery efficiency of dilution method was 56.02% and that of dialysis method was 43.15%. Our results provide necessary data for further study  on development of recombinant leptin products.

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Published

06-08-2014

How to Cite

Huong Xuan, L. M., Phuong Thao, D. T., & Thuoc, T. L. (2014). A study on refolding recombinant human leptin protein from Escherichia coli inclusion body. Academia Journal of Biology, 36(1se), 54–61. https://doi.org/10.15625/0866-7160/v36n1se.4365

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