Effects of Culture Conditions of Immobilized Recombinant Escherichia coli on Cyclodextrin Glucanotransferase (CGTase) Excretion and Cell Stability

The targeting of recombinant proteins for excretion into culture medium presents significant advantages over cytoplasmic expression. However, during the excretion of recombinant protein, caution must be taken in order to avoid cell lysis due to pressure build-up through overproduction of the express...

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Main Authors:	Rohaida, Che Man, Ahmad Fauzi, Ismail, Siti Fatima Zaharah, Mohd Fuzi, Nazlee Faisal, Ghazali, Rosli, Md. Illias
Format:	Article
Language:	English
Published:	Elsevier Ltd 2016
Subjects:	TP Chemical technology
Online Access:	http://umpir.ump.edu.my/id/eprint/11802/ http://umpir.ump.edu.my/id/eprint/11802/ http://umpir.ump.edu.my/id/eprint/11802/ http://umpir.ump.edu.my/id/eprint/11802/1/Effects%20of%20Culture%20Conditions%20of%20Immobilized%20Recombinant%20Escherichia%20coli%20on%20Cyclodextrin%20Glucanotransferase%20%28CGTase%29%20Excretion%20and%20Cell%20Stability.pdf

id	ump-11802
recordtype	eprints
spelling	ump-118022017-10-30T01:59:20Z http://umpir.ump.edu.my/id/eprint/11802/ Effects of Culture Conditions of Immobilized Recombinant Escherichia coli on Cyclodextrin Glucanotransferase (CGTase) Excretion and Cell Stability Rohaida, Che Man Ahmad Fauzi, Ismail Siti Fatima Zaharah, Mohd Fuzi Nazlee Faisal, Ghazali Rosli, Md. Illias TP Chemical technology The targeting of recombinant proteins for excretion into culture medium presents significant advantages over cytoplasmic expression. However, during the excretion of recombinant protein, caution must be taken in order to avoid cell lysis due to pressure build-up through overproduction of the expressed recombinant protein in the periplasmic space. In the present study, recombinant Escherichia coli expressing cyclodextrin glucanotransferase (CGTase) was immobilized by adsorption and entrapment in a porous hollow fiber membrane. The effects of culture conditions (post induction time, agitation rate and pH) on CGTase excretion, cell lysis and plasmid stability of immobilized cells were studied. The optimum post induction time, agitation rate and pH were found to be 24 h, 200 rpm and pH 9, respectively. The immobilized cells exhibited a 2.8–4.6-fold increase in CGTase excretion, a 16-95% reduction of cell lysis and a 323-464% increase in plasmid stability compared with free cells. Hence, immobilizing E. coli using a porous hollow fiber membrane proved to be valuable for the excretion of a recombinant protein and increased cell viability. Elsevier Ltd 2016 Article PeerReviewed application/pdf en http://umpir.ump.edu.my/id/eprint/11802/1/Effects%20of%20Culture%20Conditions%20of%20Immobilized%20Recombinant%20Escherichia%20coli%20on%20Cyclodextrin%20Glucanotransferase%20%28CGTase%29%20Excretion%20and%20Cell%20Stability.pdf Rohaida, Che Man and Ahmad Fauzi, Ismail and Siti Fatima Zaharah, Mohd Fuzi and Nazlee Faisal, Ghazali and Rosli, Md. Illias (2016) Effects of Culture Conditions of Immobilized Recombinant Escherichia coli on Cyclodextrin Glucanotransferase (CGTase) Excretion and Cell Stability. Process Biochemistry, 51 (4). pp. 474-483. ISSN 1359-5113 http://dx.doi.org/10.1016/j.procbio.2016.01.002 DOI: 10.1016/j.procbio.2016.01.002
repository_type	Digital Repository
institution_category	Local University
institution	Universiti Malaysia Pahang
building	UMP Institutional Repository
collection	Online Access
language	English
topic	TP Chemical technology
spellingShingle	TP Chemical technology Rohaida, Che Man Ahmad Fauzi, Ismail Siti Fatima Zaharah, Mohd Fuzi Nazlee Faisal, Ghazali Rosli, Md. Illias Effects of Culture Conditions of Immobilized Recombinant Escherichia coli on Cyclodextrin Glucanotransferase (CGTase) Excretion and Cell Stability
description	The targeting of recombinant proteins for excretion into culture medium presents significant advantages over cytoplasmic expression. However, during the excretion of recombinant protein, caution must be taken in order to avoid cell lysis due to pressure build-up through overproduction of the expressed recombinant protein in the periplasmic space. In the present study, recombinant Escherichia coli expressing cyclodextrin glucanotransferase (CGTase) was immobilized by adsorption and entrapment in a porous hollow fiber membrane. The effects of culture conditions (post induction time, agitation rate and pH) on CGTase excretion, cell lysis and plasmid stability of immobilized cells were studied. The optimum post induction time, agitation rate and pH were found to be 24 h, 200 rpm and pH 9, respectively. The immobilized cells exhibited a 2.8–4.6-fold increase in CGTase excretion, a 16-95% reduction of cell lysis and a 323-464% increase in plasmid stability compared with free cells. Hence, immobilizing E. coli using a porous hollow fiber membrane proved to be valuable for the excretion of a recombinant protein and increased cell viability.
format	Article
author	Rohaida, Che Man Ahmad Fauzi, Ismail Siti Fatima Zaharah, Mohd Fuzi Nazlee Faisal, Ghazali Rosli, Md. Illias
author_facet	Rohaida, Che Man Ahmad Fauzi, Ismail Siti Fatima Zaharah, Mohd Fuzi Nazlee Faisal, Ghazali Rosli, Md. Illias
author_sort	Rohaida, Che Man
title	Effects of Culture Conditions of Immobilized Recombinant Escherichia coli on Cyclodextrin Glucanotransferase (CGTase) Excretion and Cell Stability
title_short	Effects of Culture Conditions of Immobilized Recombinant Escherichia coli on Cyclodextrin Glucanotransferase (CGTase) Excretion and Cell Stability
title_full	Effects of Culture Conditions of Immobilized Recombinant Escherichia coli on Cyclodextrin Glucanotransferase (CGTase) Excretion and Cell Stability
title_fullStr	Effects of Culture Conditions of Immobilized Recombinant Escherichia coli on Cyclodextrin Glucanotransferase (CGTase) Excretion and Cell Stability
title_full_unstemmed	Effects of Culture Conditions of Immobilized Recombinant Escherichia coli on Cyclodextrin Glucanotransferase (CGTase) Excretion and Cell Stability
title_sort	effects of culture conditions of immobilized recombinant escherichia coli on cyclodextrin glucanotransferase (cgtase) excretion and cell stability
publisher	Elsevier Ltd
publishDate	2016
url	http://umpir.ump.edu.my/id/eprint/11802/ http://umpir.ump.edu.my/id/eprint/11802/ http://umpir.ump.edu.my/id/eprint/11802/ http://umpir.ump.edu.my/id/eprint/11802/1/Effects%20of%20Culture%20Conditions%20of%20Immobilized%20Recombinant%20Escherichia%20coli%20on%20Cyclodextrin%20Glucanotransferase%20%28CGTase%29%20Excretion%20and%20Cell%20Stability.pdf
first_indexed	2023-09-18T22:12:47Z
last_indexed	2023-09-18T22:12:47Z
_version_	1777415138485207040

Effects of Culture Conditions of Immobilized Recombinant Escherichia coli on Cyclodextrin Glucanotransferase (CGTase) Excretion and Cell Stability

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