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Production of recombinant human epidermal growth factor in Pichia pastoris

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 m...

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Autores principales: Eissazadeh, Samira, Moeini, Hassan, Dezfouli, Maryam Ghandizadeh, Heidary, Somayyeh, Nelofer, Rubina, Abdullah, Mohd Puad
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5470459/
https://www.ncbi.nlm.nih.gov/pubmed/27998673
http://dx.doi.org/10.1016/j.bjm.2016.10.017
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author Eissazadeh, Samira
Moeini, Hassan
Dezfouli, Maryam Ghandizadeh
Heidary, Somayyeh
Nelofer, Rubina
Abdullah, Mohd Puad
author_facet Eissazadeh, Samira
Moeini, Hassan
Dezfouli, Maryam Ghandizadeh
Heidary, Somayyeh
Nelofer, Rubina
Abdullah, Mohd Puad
author_sort Eissazadeh, Samira
collection PubMed
description 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 2.27 μg/mL was obtained through an induction of the culture with 0.5% (v/v) methanol for 60 h. 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.
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spelling pubmed-54704592017-06-23 Production of recombinant human epidermal growth factor in Pichia pastoris Eissazadeh, Samira Moeini, Hassan Dezfouli, Maryam Ghandizadeh Heidary, Somayyeh Nelofer, Rubina Abdullah, Mohd Puad Braz J Microbiol Biotechnology and Industrial Microbiology 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 2.27 μg/mL was obtained through an induction of the culture with 0.5% (v/v) methanol for 60 h. 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. Elsevier 2017-04-07 /pmc/articles/PMC5470459/ /pubmed/27998673 http://dx.doi.org/10.1016/j.bjm.2016.10.017 Text en © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Biotechnology and Industrial Microbiology
Eissazadeh, Samira
Moeini, Hassan
Dezfouli, Maryam Ghandizadeh
Heidary, Somayyeh
Nelofer, Rubina
Abdullah, Mohd Puad
Production of recombinant human epidermal growth factor in Pichia pastoris
title Production of recombinant human epidermal growth factor in Pichia pastoris
title_full Production of recombinant human epidermal growth factor in Pichia pastoris
title_fullStr Production of recombinant human epidermal growth factor in Pichia pastoris
title_full_unstemmed Production of recombinant human epidermal growth factor in Pichia pastoris
title_short Production of recombinant human epidermal growth factor in Pichia pastoris
title_sort production of recombinant human epidermal growth factor in pichia pastoris
topic Biotechnology and Industrial Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5470459/
https://www.ncbi.nlm.nih.gov/pubmed/27998673
http://dx.doi.org/10.1016/j.bjm.2016.10.017
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