Improved Production of Recombinant Human β-NGF in Escherichia coli – a Bioreactor Scale Study

Human nerve growth factor β (β-NGF) is considered a major therapeutic agent for treatment of neurodegenerative diseases. We have previously reported the optimized conditions for β-NGF overproduction in Escherichia coli in a shake-flask culture. In this study the optimal %DO (dissolved oxygen) and po...

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Main Authors: POURIA GHOLAMI TILKO, SEYEDEH MAHDIEH SADAT, ZAHRA HAJIHASSAN
Format: Article
Language:English
Published: Sciendo 2018-09-01
Series:Polish Journal of Microbiology
Subjects:
Online Access:https://www.exeley.com/exeley/journals/polish_journal_of_microbiology/67/3/pdf/10.21307_pjm-2018-045.pdf
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author POURIA GHOLAMI TILKO
SEYEDEH MAHDIEH SADAT
ZAHRA HAJIHASSAN
author_facet POURIA GHOLAMI TILKO
SEYEDEH MAHDIEH SADAT
ZAHRA HAJIHASSAN
author_sort POURIA GHOLAMI TILKO
collection DOAJ
description Human nerve growth factor β (β-NGF) is considered a major therapeutic agent for treatment of neurodegenerative diseases. We have previously reported the optimized conditions for β-NGF overproduction in Escherichia coli in a shake-flask culture. In this study the optimal %DO (dissolved oxygen) and post induction temperature values for improved production of β-NGF were found in the bioreactor scale using response surface methodology (RSM) as the most common statistical method. Also, for further enhancement of the yield, different post-induction periods of time were selected for testing. In all experiments, the productivity level and bacterial cell growth were evaluated by western blotting technique and monitoring of absorbance at 600 nm, respectively. Our results indicated that %DO, the post-induction time and temperature have significant effects on the production of β-NGF. After 2 hours of induction, the low post induction temperature of 32°C and 20% DO were used to increase the production of β-NGF in a 5-l bioreactor. Another important result obtained in this study was that the improved β-NGF production was not achieved at highest dry cell weigh or highest cell growth. These results are definitely of importance for industrial β-NGF production.
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spelling doaj.art-624bd498b4744df49a414d5f498cb37d2022-12-22T03:41:42ZengSciendoPolish Journal of Microbiology1733-13312544-46462018-09-0167310.21307/pjm-2018-045Improved Production of Recombinant Human β-NGF in Escherichia coli – a Bioreactor Scale StudyPOURIA GHOLAMI TILKO0SEYEDEH MAHDIEH SADAT1ZAHRA HAJIHASSAN2Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran,Tehran,IranDepartment of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran,Tehran,IranDepartment of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran,Tehran,IranHuman nerve growth factor β (β-NGF) is considered a major therapeutic agent for treatment of neurodegenerative diseases. We have previously reported the optimized conditions for β-NGF overproduction in Escherichia coli in a shake-flask culture. In this study the optimal %DO (dissolved oxygen) and post induction temperature values for improved production of β-NGF were found in the bioreactor scale using response surface methodology (RSM) as the most common statistical method. Also, for further enhancement of the yield, different post-induction periods of time were selected for testing. In all experiments, the productivity level and bacterial cell growth were evaluated by western blotting technique and monitoring of absorbance at 600 nm, respectively. Our results indicated that %DO, the post-induction time and temperature have significant effects on the production of β-NGF. After 2 hours of induction, the low post induction temperature of 32°C and 20% DO were used to increase the production of β-NGF in a 5-l bioreactor. Another important result obtained in this study was that the improved β-NGF production was not achieved at highest dry cell weigh or highest cell growth. These results are definitely of importance for industrial β-NGF production.https://www.exeley.com/exeley/journals/polish_journal_of_microbiology/67/3/pdf/10.21307_pjm-2018-045.pdfbioreactorRSMβ-NGF over productionE. coli
spellingShingle POURIA GHOLAMI TILKO
SEYEDEH MAHDIEH SADAT
ZAHRA HAJIHASSAN
Improved Production of Recombinant Human β-NGF in Escherichia coli – a Bioreactor Scale Study
Polish Journal of Microbiology
bioreactor
RSM
β-NGF over production
E. coli
title Improved Production of Recombinant Human β-NGF in Escherichia coli – a Bioreactor Scale Study
title_full Improved Production of Recombinant Human β-NGF in Escherichia coli – a Bioreactor Scale Study
title_fullStr Improved Production of Recombinant Human β-NGF in Escherichia coli – a Bioreactor Scale Study
title_full_unstemmed Improved Production of Recombinant Human β-NGF in Escherichia coli – a Bioreactor Scale Study
title_short Improved Production of Recombinant Human β-NGF in Escherichia coli – a Bioreactor Scale Study
title_sort improved production of recombinant human β ngf in escherichia coli a bioreactor scale study
topic bioreactor
RSM
β-NGF over production
E. coli
url https://www.exeley.com/exeley/journals/polish_journal_of_microbiology/67/3/pdf/10.21307_pjm-2018-045.pdf
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