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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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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format | Article |
id | utm.eprints-69325 |
institution | Universiti Teknologi Malaysia - ePrints |
last_indexed | 2024-03-05T20:00:07Z |
publishDate | 2016 |
publisher | Elsevier Ltd |
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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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