SCREENING THE EFFECT OF THE EXPRESSION MEDIUM AND GROWTH CONDITIONS ON THE PERFORMANCE OF ENGINEERED XYLANASE PRODUCED BY IMMOBILIZED RECOMBINANT E. COLI
Escherichia coli is the most prevalent host organism for the production of recombinant enzymes. This was feasible due to the possibility of genetic modification and the availability of multiple E. coli strains as recombinant systems. The primary disadvantage of using E. coli as a host, however, is...
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| Format: | Article |
| Language: | English |
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2023
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| Online Access: | http://eprints.uthm.edu.my/10120/1/J16259_bbcb103dab1c86792ecac69a93328e4c.pdf |
| _version_ | 1825638294086483968 |
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| author | Mohd Fuzi, Siti Fatimah Zaharah Nor Ashikin, Nur Atiqah Lyana Low Kheng Oon, Low Kheng Oon |
| author_facet | Mohd Fuzi, Siti Fatimah Zaharah Nor Ashikin, Nur Atiqah Lyana Low Kheng Oon, Low Kheng Oon |
| author_sort | Mohd Fuzi, Siti Fatimah Zaharah |
| collection | UTHM |
| description | Escherichia coli is the most prevalent host organism for the production of recombinant enzymes. This was feasible due to the possibility of genetic modification and the availability of
multiple E. coli strains as recombinant systems. The primary disadvantage of using E. coli as a host, however, is bacterial cell lysis due to tension build-up in the periplasmic space caused by the overexpression of the recombinant enzyme. Therefore, immobilization is preferable to cytoplasmic excretion for directing the expression of recombinant enzymes into the culture medium. This research investigated the effect of graphene oxide (GO) on the xylanase and
β-galactosidase activity of immobilized recombinant E. coli. The effect of culture conditions (expression medium, IPTG, post induction temperature, post induction duration, agitation
rate, and pH) on xylanase excretion and cell survival of an immobilized cell was studied using the one factor at a time (OFAT) method. After 24 hours of induction, using terrific broth (TB) as a medium increased xylanase excretion to 0.060 U/ml and resulted in decreased βgalactosidase activity (1.218 U/ml). Apart from that, a lower concentration of isopropyl -D-1- thiogalactopyranoside (IPTG) at 0.01 mM, a lower post-induction temperature (25°C), a 5- hour post-induction time, neutral pH, and 150 rpm significantly increased the xylanase
excretion of immobilized cells with low β-galactosidase activity. This study established that immobilizing recombinant E. coli on GO may be advantageous for the excretion of
recombinant proteins with a high cell viability |
| first_indexed | 2024-03-05T22:04:36Z |
| format | Article |
| id | uthm.eprints-10120 |
| institution | Universiti Tun Hussein Onn Malaysia |
| language | English |
| last_indexed | 2024-03-05T22:04:36Z |
| publishDate | 2023 |
| record_format | dspace |
| spelling | uthm.eprints-101202023-10-17T07:25:56Z http://eprints.uthm.edu.my/10120/ SCREENING THE EFFECT OF THE EXPRESSION MEDIUM AND GROWTH CONDITIONS ON THE PERFORMANCE OF ENGINEERED XYLANASE PRODUCED BY IMMOBILIZED RECOMBINANT E. COLI Mohd Fuzi, Siti Fatimah Zaharah Nor Ashikin, Nur Atiqah Lyana Low Kheng Oon, Low Kheng Oon R Medicine (General) Escherichia coli is the most prevalent host organism for the production of recombinant enzymes. This was feasible due to the possibility of genetic modification and the availability of multiple E. coli strains as recombinant systems. The primary disadvantage of using E. coli as a host, however, is bacterial cell lysis due to tension build-up in the periplasmic space caused by the overexpression of the recombinant enzyme. Therefore, immobilization is preferable to cytoplasmic excretion for directing the expression of recombinant enzymes into the culture medium. This research investigated the effect of graphene oxide (GO) on the xylanase and β-galactosidase activity of immobilized recombinant E. coli. The effect of culture conditions (expression medium, IPTG, post induction temperature, post induction duration, agitation rate, and pH) on xylanase excretion and cell survival of an immobilized cell was studied using the one factor at a time (OFAT) method. After 24 hours of induction, using terrific broth (TB) as a medium increased xylanase excretion to 0.060 U/ml and resulted in decreased βgalactosidase activity (1.218 U/ml). Apart from that, a lower concentration of isopropyl -D-1- thiogalactopyranoside (IPTG) at 0.01 mM, a lower post-induction temperature (25°C), a 5- hour post-induction time, neutral pH, and 150 rpm significantly increased the xylanase excretion of immobilized cells with low β-galactosidase activity. This study established that immobilizing recombinant E. coli on GO may be advantageous for the excretion of recombinant proteins with a high cell viability 2023 Article PeerReviewed text en http://eprints.uthm.edu.my/10120/1/J16259_bbcb103dab1c86792ecac69a93328e4c.pdf Mohd Fuzi, Siti Fatimah Zaharah and Nor Ashikin, Nur Atiqah Lyana and Low Kheng Oon, Low Kheng Oon (2023) SCREENING THE EFFECT OF THE EXPRESSION MEDIUM AND GROWTH CONDITIONS ON THE PERFORMANCE OF ENGINEERED XYLANASE PRODUCED BY IMMOBILIZED RECOMBINANT E. COLI. Jurnal Teknologi, 85 (3). pp. 183-193. ISSN 2180–3722 https://doi.org/10.11113/jurnalteknologi.v85.18143%7C |
| spellingShingle | R Medicine (General) Mohd Fuzi, Siti Fatimah Zaharah Nor Ashikin, Nur Atiqah Lyana Low Kheng Oon, Low Kheng Oon SCREENING THE EFFECT OF THE EXPRESSION MEDIUM AND GROWTH CONDITIONS ON THE PERFORMANCE OF ENGINEERED XYLANASE PRODUCED BY IMMOBILIZED RECOMBINANT E. COLI |
| title | SCREENING THE EFFECT OF THE EXPRESSION MEDIUM AND GROWTH CONDITIONS ON THE PERFORMANCE OF ENGINEERED XYLANASE PRODUCED BY IMMOBILIZED RECOMBINANT E. COLI |
| title_full | SCREENING THE EFFECT OF THE EXPRESSION MEDIUM AND GROWTH CONDITIONS ON THE PERFORMANCE OF ENGINEERED XYLANASE PRODUCED BY IMMOBILIZED RECOMBINANT E. COLI |
| title_fullStr | SCREENING THE EFFECT OF THE EXPRESSION MEDIUM AND GROWTH CONDITIONS ON THE PERFORMANCE OF ENGINEERED XYLANASE PRODUCED BY IMMOBILIZED RECOMBINANT E. COLI |
| title_full_unstemmed | SCREENING THE EFFECT OF THE EXPRESSION MEDIUM AND GROWTH CONDITIONS ON THE PERFORMANCE OF ENGINEERED XYLANASE PRODUCED BY IMMOBILIZED RECOMBINANT E. COLI |
| title_short | SCREENING THE EFFECT OF THE EXPRESSION MEDIUM AND GROWTH CONDITIONS ON THE PERFORMANCE OF ENGINEERED XYLANASE PRODUCED BY IMMOBILIZED RECOMBINANT E. COLI |
| title_sort | screening the effect of the expression medium and growth conditions on the performance of engineered xylanase produced by immobilized recombinant e coli |
| topic | R Medicine (General) |
| url | http://eprints.uthm.edu.my/10120/1/J16259_bbcb103dab1c86792ecac69a93328e4c.pdf |
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