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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 advan-tages 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 expres...

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Main Authors: Mohd Fuzi/, Siti Fatimah Zaharah, Che Man, Rohaida, Ismail, Ahmad Fauzi, Ghazali, Nazlee Faisal, Md Illias, Rosli
Format: Article
Language:English
Published: Elsevier Ltd. 2016
Subjects:
QD Chemistry
Online Access:http://eprints.uthm.edu.my/4189/1/AJ%202016%20%2823%29.pdf
http://eprints.uthm.edu.my/4189/
http://dx.doi.org/10.1016/j.procbio.2016.01.002
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spelling my.uthm.eprints.41892021-11-28T06:53:04Z http://eprints.uthm.edu.my/4189/ Effects of culture conditions of immobilized recombinant Escherichia coli on cyclodextrin glucanotransferase (CGTase) excretion and cell stability Mohd Fuzi/, Siti Fatimah Zaharah Che Man, Rohaida Ismail, Ahmad Fauzi Ghazali, Nazlee Faisal Md Illias, Rosli QD Chemistry The targeting of recombinant proteins for excretion into culture medium presents significant advan-tages 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 express-ing 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 immo-bilized 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 text en http://eprints.uthm.edu.my/4189/1/AJ%202016%20%2823%29.pdf Mohd Fuzi/, Siti Fatimah Zaharah and Che Man, Rohaida and Ismail, Ahmad Fauzi and Ghazali, Nazlee Faisal and Md Illias, Rosli (2016) Effects of culture conditions of immobilized recombinant Escherichia coli on cyclodextrin glucanotransferase (CGTase) excretion and cell stability. Process Biochemistry, 51. pp. 474-483. ISSN 1359-5113 http://dx.doi.org/10.1016/j.procbio.2016.01.002
institution Universiti Tun Hussein Onn Malaysia
building UTHM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Tun Hussein Onn Malaysia
content_source UTHM Institutional Repository
url_provider http://eprints.uthm.edu.my/
language English
topic QD Chemistry
spellingShingle QD Chemistry
Mohd Fuzi/, Siti Fatimah Zaharah
Che Man, Rohaida
Ismail, Ahmad Fauzi
Ghazali, Nazlee Faisal
Md Illias, Rosli
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 advan-tages 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 express-ing 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 immo-bilized 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 Mohd Fuzi/, Siti Fatimah Zaharah
Che Man, Rohaida
Ismail, Ahmad Fauzi
Ghazali, Nazlee Faisal
Md Illias, Rosli
author_facet Mohd Fuzi/, Siti Fatimah Zaharah
Che Man, Rohaida
Ismail, Ahmad Fauzi
Ghazali, Nazlee Faisal
Md Illias, Rosli
author_sort Mohd Fuzi/, Siti Fatimah Zaharah
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://eprints.uthm.edu.my/4189/1/AJ%202016%20%2823%29.pdf
http://eprints.uthm.edu.my/4189/
http://dx.doi.org/10.1016/j.procbio.2016.01.002
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score 13.211869

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