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: Che Man, Rohaida, Ismail, Ahmad Fauzi, Mohd. Fuzi, Siti Fatimah Zaharah, Ghazali, Nazlee Faisal, Md. Illias, Rosli
Format: Article
Published: Elsevier Ltd 2016
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author Che Man, Rohaida
Ismail, Ahmad Fauzi
Mohd. Fuzi, Siti Fatimah Zaharah
Ghazali, Nazlee Faisal
Md. Illias, Rosli
author_facet Che Man, Rohaida
Ismail, Ahmad Fauzi
Mohd. Fuzi, Siti Fatimah Zaharah
Ghazali, Nazlee Faisal
Md. Illias, Rosli
author_sort Che Man, Rohaida
collection ePrints
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 24h, 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.
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spelling utm.eprints-693252017-11-22T00:45:09Z http://eprints.utm.my/69325/ Effects of culture conditions of immobilized recombinant Escherichia coli on cyclodextrin glucanotransferase (CGTase) excretion and cell stability Che Man, Rohaida Ismail, Ahmad Fauzi Mohd. Fuzi, Siti Fatimah Zaharah Ghazali, Nazlee Faisal Md. Illias, Rosli 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 24h, 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 Che Man, Rohaida and Ismail, Ahmad Fauzi and Mohd. Fuzi, Siti Fatimah Zaharah 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 (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
spellingShingle TP Chemical technology
Che Man, Rohaida
Ismail, Ahmad Fauzi
Mohd. Fuzi, Siti Fatimah Zaharah
Ghazali, Nazlee Faisal
Md. Illias, Rosli
Effects of culture conditions of immobilized recombinant Escherichia coli on cyclodextrin glucanotransferase (CGTase) excretion and cell stability
title 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_short 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
topic TP Chemical technology
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