Effects of culture conditions of immobilized recombinant Escherichia coli on cyclodextrin glucanotransferase (CGTase) excretionand 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 mustbe taken in order to avoid cell lysis due to pressure build-up through overproduction of the express...
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| Format: | Article |
| Language: | English |
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Elsevier
2016
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| Online Access: | http://eprints.uthm.edu.my/5258/ http://eprints.uthm.edu.my/5258/1/AJ%202016%20%2872%29.pdf |
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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 |
| building | UTHM Institutional Repository |
| collection | Online Access |
| 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 mustbe taken in order to avoid cell lysis due to pressure build-up through overproduction of the expressedrecombinant 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 poroushollow fiber membrane. The effects of culture conditions (post induction time, agitation rate and pH) onCGTase excretion, cell lysis and plasmid stability of immobilized cells were studied. The optimum postinduction 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 anda 323–464% increase in plasmid stability compared with free cells. Hence, immobilizing E. coli using aporous hollow fiber membrane proved to be valuable for the excretion of a recombinant protein andincreased cell viability. |
| first_indexed | 2025-11-15T20:11:21Z |
| format | Article |
| id | uthm-5258 |
| institution | Universiti Tun Hussein Onn Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T20:11:21Z |
| publishDate | 2016 |
| publisher | Elsevier |
| recordtype | eprints |
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| spelling | uthm-52582022-01-06T08:11:16Z http://eprints.uthm.edu.my/5258/ Effects of culture conditions of immobilized recombinant Escherichia coli on cyclodextrin glucanotransferase (CGTase) excretionand cell stability Che Man, Rohaida Ismail, Ahmad Fauzi Mohd Fuzi, Siti Fatimah Zaharah 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 mustbe taken in order to avoid cell lysis due to pressure build-up through overproduction of the expressedrecombinant 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 poroushollow fiber membrane. The effects of culture conditions (post induction time, agitation rate and pH) onCGTase excretion, cell lysis and plasmid stability of immobilized cells were studied. The optimum postinduction 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 anda 323–464% increase in plasmid stability compared with free cells. Hence, immobilizing E. coli using aporous hollow fiber membrane proved to be valuable for the excretion of a recombinant protein andincreased cell viability. Elsevier 2016 Article PeerReviewed text en http://eprints.uthm.edu.my/5258/1/AJ%202016%20%2872%29.pdf 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) excretionand cell stability. Process Biochemistry, 51. pp. 474-483. ISSN 1359-5113 http://dx.doi.org/10.1016/j.procbio.2016.01.002 |
| spellingShingle | QD Chemistry 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) excretionand cell stability |
| title | Effects of culture conditions of immobilized recombinant Escherichia coli on cyclodextrin glucanotransferase (CGTase) excretionand cell stability |
| title_full | Effects of culture conditions of immobilized recombinant Escherichia coli on cyclodextrin glucanotransferase (CGTase) excretionand cell stability |
| title_fullStr | Effects of culture conditions of immobilized recombinant Escherichia coli on cyclodextrin glucanotransferase (CGTase) excretionand cell stability |
| title_full_unstemmed | Effects of culture conditions of immobilized recombinant Escherichia coli on cyclodextrin glucanotransferase (CGTase) excretionand cell stability |
| title_short | Effects of culture conditions of immobilized recombinant Escherichia coli on cyclodextrin glucanotransferase (CGTase) excretionand cell stability |
| title_sort | effects of culture conditions of immobilized recombinant escherichia coli on cyclodextrin glucanotransferase (cgtase) excretionand cell stability |
| topic | QD Chemistry |
| url | http://eprints.uthm.edu.my/5258/ http://eprints.uthm.edu.my/5258/ http://eprints.uthm.edu.my/5258/1/AJ%202016%20%2872%29.pdf |