Optimization extracellular secretion of Recombinant Human Epidermal Growth Factor (hEGF) in Escherichia coli BL21 (DE3) pD881-OmpA-hEGF by using Response Surface Method (RSM)

  • Annisa Indriyani Biomolecular Laboratory of Health and Food, Departement of Chemistry, Faculty of Mathematics and Natural Science, Padjadjaran University, Indonesia, +62227794391
  • Nenden Indriyani Anggraeni Biomolecular Laboratory of Health and Food, Departement of Chemistry, Faculty of Mathematics and Natural Science, Padjadjaran University, Indonesia, +62227794391
  • Sriwidodo Department of Pharmaceutics & Pharmaceutical Technology, Faculty of Pharmacy, Padjadjaran University, Indonesia, +62227794391
  • Iman Permana Maksum Biomolecular Laboratory of Health and Food, Departement of Chemistry, Faculty of Mathematics and Natural Science, Padjadjaran University, Indonesia, +62227794391

Abstract

Human Epidermal Growth Factor is techno-economically protein that has the function for stimulates a process of proliferation and cell differentiation. Because of that function, it can be a candidate for wound healing in ulcer diabetic. In the previous study, hEGF can extracellular secreted by OmpA signal peptide using recombinant DNA. The production method of recombinant hEGF will be extracellular secreted by Escherichia coli. However, the optimum secretion of OmpA has not been studied. Therefore the purpose of this study is to determine the optimum conditions using the RSM method. This research begins with a rejuvenation of culture, extracellular secretion, and optimization with the RSM method, followed by protein production at optimum condition. Secreted recombinant protein measured by Sandwich ELISA method and LOWRY. The result showed that the optimum condition was found in the medium concentration of 1.5x and induction time at 2 hours 10 minutes and the total protein yield of expression was 21.247 mg/mL with optimization percentage 2.96% and efficiency 50.56 % and recombinant protein hEGF concentration in fermentation media 416 μg/mL

Keywords: extracellular secretion, LOWRY, recombinant, RSM and Sandwich ELISA

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Published
2019-07-12
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How to Cite
Annisa Indriyani, Nenden Indriyani Anggraeni, Sriwidodo, & Iman Permana Maksum. (2019). Optimization extracellular secretion of Recombinant Human Epidermal Growth Factor (hEGF) in Escherichia coli BL21 (DE3) pD881-OmpA-hEGF by using Response Surface Method (RSM). International Journal of Research in Pharmaceutical Sciences, 10(3), 1824-1831. https://doi.org/10.26452/ijrps.v10i3.1378
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Original Articles
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