COMPARISON OF EXTRACELLULAR SECRETION OF RECOMBINANT HUMAN EPIDERMAL GROWTH FACTOR USING TORA AND PELB SIGNAL PEPTIDES IN ESCHERICHIA COLI BL21 (DE3)

  • RIMA MELATI Biomolecular Laboratory of Health and Food, Department of Chemistry, Faculty of Mathematics and Natural Science, Padjadjaran University, Indonesia.
  • ANNISA INDRIYANI Biomolecular Laboratory of Health and Food, Department of Chemistry, Faculty of Mathematics and Natural Science, Padjadjaran University, Indonesia.
  • SHABARNI GAFFAR Biomolecular Laboratory of Health and Food, Department of Chemistry, Faculty of Mathematics and Natural Science, Padjadjaran University, Indonesia.
  • SRIWIDODO Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, Padjadjaran University, Indonesia.
  • IMAN PERMANA MAKSUM Biomolecular Laboratory of Health and Food, Department of Chemistry, Faculty of Mathematics and Natural Science, Padjadjaran University, Indonesia.

Abstract

Objective: The objective of this study was to evaluate two signal peptides (TorA and PelB), representing the most common secretion pathways in Escherichia coli, for their ability to secrete recombinant human epidermal growth factor (rhEGF) protein in the extracellular expression.


Methods: E. coli BL21 (DE3) as the host cell to be transformed using recombinant plasmid pD881-TorA the consensus already containing hEGF gene and the signal peptide TorA or PelB, then expressed by L-rhamnose induction. rhEGF purified by heat treatment and ion-exchange chromatography. The hEGF protein was characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and ELISA.


Results: The result showed that PelB was secreting more hEGF protein compared to TorA with protein expression results of 48.2 μg/L and purification results of 0.360 μg/L, with a purity level of 83%.


Conclusion: The results of this study explain in extracellular expression of hEGF protein in E. coli, PelB helps hEGF protein secretion to culture media better than TorA.

Keywords: Human epidermal growth factor, Escherichia coli, Extracellular expression, TorA, PelB

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RIMA MELATI, ANNISA INDRIYANI, SHABARNI GAFFAR, SRIWIDODO, and IMAN PERMANA MAKSUM. “COMPARISON OF EXTRACELLULAR SECRETION OF RECOMBINANT HUMAN EPIDERMAL GROWTH FACTOR USING TORA AND PELB SIGNAL PEPTIDES IN ESCHERICHIA COLI BL21 (DE3)”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 12, no. 11, Sept. 2019, pp. 81-84, doi:10.22159/ajpcr.2019.v12i11.35316.
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