Optimization of Secreted Recombinant Human Epidermal Growth Factor Production Using Pectate Lyase B from Escherichia Coli BL21(DE3) by Central Composite Design and Its Production in High Cell Density Culture

Journal of Pharmacy & Bioallied Sciences
Sriwidodo SriwidodoIndah U Putri

Abstract

Human Epidermal Growth Factor (hEGF) is a potential therapeutic protein that has been widely used as a healing agent for various chronic wounds. It induces the proliferation and metabolism of epithelial cells, regenerates skin cells, and validates skin elasticity. In the previous study, recombinant hEGF (rhEGF) had been successfully expressed extracellularly in Escherichia coli (E. coli) BL21 (DE3) using pectate lyase B (PelB) signal peptide. The previous study has shown that the medium concentration and the induction time influenced the production of rhEGF. Therefore, this study was conducted to optimize the induction time and medium concentration for rhEGF extracellular secretion then followed by scale-up production. This experiment was carried out using E. coli BL21 (DE3) which contains pD881 plasmid that carries hEGF and PelB gene. Optimization design of induction time and medium concentration were obtained using Central Composite Design (CCD). The method of study started by the rejuvenation of E. coli culture, extracellular secretion, and optimization in the flask scale then followed by scaled-up production with high-cell density culture in the fermenter. The optimization was carried out using Response Surface Methodology ...Continue Reading

References

Jan 1, 1979·Annual Review of Biochemistry·G Carpenter, S Cohen
Sep 30, 2009·Journal of Molecular Biology·Haeyoung JeongJihyun F Kim
Dec 22, 2016·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Samira EissazadehMohd Puad Abdullah

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Citations

Oct 17, 2020·Journal of Experimental Pharmacology·Sriwidodo SriwidodoAbd Kakhar Umar

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Methods Mentioned

BETA
electrophoresis
enzyme-linked immunosorbent assay
ELISA

Software Mentioned

Minitab

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