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...
Main Authors: | , , |
---|---|
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 |
_version_ | 1811220670098440192 |
---|---|
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. |
first_indexed | 2024-04-12T07:46:35Z |
format | Article |
id | doaj.art-624bd498b4744df49a414d5f498cb37d |
institution | Directory Open Access Journal |
issn | 1733-1331 2544-4646 |
language | English |
last_indexed | 2024-04-12T07:46:35Z |
publishDate | 2018-09-01 |
publisher | Sciendo |
record_format | Article |
series | Polish Journal of Microbiology |
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 |
work_keys_str_mv | AT pouriagholamitilko improvedproductionofrecombinanthumanbngfinescherichiacoliabioreactorscalestudy AT seyedehmahdiehsadat improvedproductionofrecombinanthumanbngfinescherichiacoliabioreactorscalestudy AT zahrahajihassan improvedproductionofrecombinanthumanbngfinescherichiacoliabioreactorscalestudy |