Effects of culture conditions of immobilized recombinant Escherichia coli on cyclodextrin glucanotransferase (CGTase) excretionand cell stability

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

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Abstract

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.

Item Type: Article
Uncontrolled Keywords: Immobilized cell; Free cell; CGTase excretion; Cell lysis; Plasmid stability; Porous hollow fiber membranea;
Subjects: Q Science > QD Chemistry
Divisions: Faculty of Applied Science and Technology > Department of Physics and Chemistry
Depositing User: Mrs. Nur Nadia Md. Jurimi
Date Deposited: 06 Jan 2022 08:11
Last Modified: 06 Jan 2022 08:11
URI: http://eprints.uthm.edu.my/id/eprint/5258

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