Cell rupture of recombinant escherichia coli using high pressure homogenizer

Cell rupture is one of the earlier steps in downstream processing which are required for the recovery of biological products that are located inside cells. Cells could be disrupted either by using chemicals or mechanical method. In this study, cell rupture was carried out by mechanical force using H...

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Main Authors: Ahmad Raus, Raha, Mel, Maizirwan, Mohd Abdullah, S. N., Yusoff, K.
Format: Article
Language:English
Published: Asian Network for Scientific Information 2010
Subjects:
Online Access:http://irep.iium.edu.my/1251/1/Cell_Rupture_of_Recombinant_E._coli_Using_High_Pressure_Homogenizer.pdf
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author Ahmad Raus, Raha
Mel, Maizirwan
Mohd Abdullah, S. N.
Yusoff, K.
author_facet Ahmad Raus, Raha
Mel, Maizirwan
Mohd Abdullah, S. N.
Yusoff, K.
author_sort Ahmad Raus, Raha
collection IIUM
description Cell rupture is one of the earlier steps in downstream processing which are required for the recovery of biological products that are located inside cells. Cells could be disrupted either by using chemicals or mechanical method. In this study, cell rupture was carried out by mechanical force using High Pressure Homogenizer (HPH). The aim of this study is to identify optimal conditions of HPH to disrupt the cell wall of recombinant Escherichia coli harboring nucleocapsid (NP) gene of Newcastle Disease Virus (NDV). The optimized conditions were achieved by manipulating the independent variables of HPH such as pressure, pump speed and number of cycles through an optimization process. The efficiency of the cell disruption was determined by estimating the percentage of cell rupture as well as the amount of NP protein released from the cell lysis. Through the means plot analysis of Minitab Software (Version 14.12), pressure was recognized as the main factor for achieving the highest cell rupture and the release of NP protein. The optimized conditions for obtaining the highest NP protein yield were by operating three cycles of cell rupture, homogenizer pressure of 800 bars and pump speed of 7 psi.
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spelling oai:generic.eprints.org:12512012-01-16T08:10:32Z http://irep.iium.edu.my/1251/ Cell rupture of recombinant escherichia coli using high pressure homogenizer Ahmad Raus, Raha Mel, Maizirwan Mohd Abdullah, S. N. Yusoff, K. TP248.13 Biotechnology Cell rupture is one of the earlier steps in downstream processing which are required for the recovery of biological products that are located inside cells. Cells could be disrupted either by using chemicals or mechanical method. In this study, cell rupture was carried out by mechanical force using High Pressure Homogenizer (HPH). The aim of this study is to identify optimal conditions of HPH to disrupt the cell wall of recombinant Escherichia coli harboring nucleocapsid (NP) gene of Newcastle Disease Virus (NDV). The optimized conditions were achieved by manipulating the independent variables of HPH such as pressure, pump speed and number of cycles through an optimization process. The efficiency of the cell disruption was determined by estimating the percentage of cell rupture as well as the amount of NP protein released from the cell lysis. Through the means plot analysis of Minitab Software (Version 14.12), pressure was recognized as the main factor for achieving the highest cell rupture and the release of NP protein. The optimized conditions for obtaining the highest NP protein yield were by operating three cycles of cell rupture, homogenizer pressure of 800 bars and pump speed of 7 psi. Asian Network for Scientific Information 2010 Article PeerReviewed application/pdf en http://irep.iium.edu.my/1251/1/Cell_Rupture_of_Recombinant_E._coli_Using_High_Pressure_Homogenizer.pdf Ahmad Raus, Raha and Mel, Maizirwan and Mohd Abdullah, S. N. and Yusoff, K. (2010) Cell rupture of recombinant escherichia coli using high pressure homogenizer. Journal of Applied Sciences, 10 (21). pp. 2717-2720. ISSN 1812-5662 (O), 1812-5654 (P) http://scialert.net/abstract/?doi=jas.2010.2717.2720 10.3923/jas.2010.2717.2720
spellingShingle TP248.13 Biotechnology
Ahmad Raus, Raha
Mel, Maizirwan
Mohd Abdullah, S. N.
Yusoff, K.
Cell rupture of recombinant escherichia coli using high pressure homogenizer
title Cell rupture of recombinant escherichia coli using high pressure homogenizer
title_full Cell rupture of recombinant escherichia coli using high pressure homogenizer
title_fullStr Cell rupture of recombinant escherichia coli using high pressure homogenizer
title_full_unstemmed Cell rupture of recombinant escherichia coli using high pressure homogenizer
title_short Cell rupture of recombinant escherichia coli using high pressure homogenizer
title_sort cell rupture of recombinant escherichia coli using high pressure homogenizer
topic TP248.13 Biotechnology
url http://irep.iium.edu.my/1251/1/Cell_Rupture_of_Recombinant_E._coli_Using_High_Pressure_Homogenizer.pdf
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