title: Production of recombinant human epidermal growth factor in Pichia pastoris.
abstract: This study was carried out to express human epidermal growth factor (hEGF) in Pichia pastoris GS115. For this aim, the hEGF gene was cloned into the pPIC9K expression vector, and then integrated into P. pastoris by electroporation. ELISA-based assay showed that the amount of hEGF secreted into the medium can be affected by the fermentation conditions especially by culture medium, pH and temperature. The best medium for the optimal hEGF production was BMMY buffered at a pH range of 6.0 and 7.0. The highest amount of hEGF with an average yield of 2.27μg/mL was obtained through an induction of the culture with 0.5% (v/v) methanol for 60h. The artificial neural network (ANN) analysis revealed that changes in both pH and temperature significantly affected the hEGF production with the pH change had slightly higher impact on hEGF production than variations in the temperature.
authors.affiliation: Universiti Putra Malaysia, Faculty of Biotechnology and Molecular Science, Department of Cellular and Molecular Biology, Malaysia.
authors.forename: Samira
authors.initials: S
authors.lastname: Eissazadeh
authors.affiliation: Universiti Putra Malaysia, Institute of Bioscience, Malaysia. Electronic address: moeini82@yahoo.com.
authors.forename: Hassan
authors.initials: H
authors.lastname: Moeini
authors.affiliation: Arak University, Faculty of Science, Department of Biology, Iran.
authors.forename: Maryam Ghandizadeh
authors.initials: MG
authors.lastname: Dezfouli
authors.affiliation: Universiti Putra Malaysia, Faculty of Biotechnology and Molecular Science, Department of Cellular and Molecular Biology, Malaysia.
authors.forename: Somayyeh
authors.initials: S
authors.lastname: Heidary
authors.affiliation: Pakistan Council of Scientific and Industrial Research, Laboratories Complex, Lahore, Pakistan.
authors.forename: Rubina
authors.initials: R
authors.lastname: Nelofer
authors.affiliation: Universiti Putra Malaysia, Faculty of Biotechnology and Molecular Science, Department of Cellular and Molecular Biology, Malaysia.
authors.forename: Mohd Puad
authors.initials: MP
authors.lastname: Abdullah
mesh_terms: D003001:Cloning, Molecular; D003470:Culture Media / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D004815:Epidermal Growth Factor / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D015870:Gene Expression; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor
keywords: Artificial neural network; Human epidermal growth factor; Pichia pastoris