Plasmid-based and genome-based expression of recombinant T1 lipase in sucrose-utilizing E. coli strain W
Given its thermoalkaliphilic properties, T1 lipase holds significant potential for diverse industrial applications. However, traditional expression methods in Escherichia coli, specifically the plasmid-based system, present challenges of exerting metabolic burden on host cells and elevated costs due...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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University of Malaya
2024
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| Online Access: | http://psasir.upm.edu.my/id/eprint/112021/1/112021.pdf |
| _version_ | 1825940057909886976 |
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| author | Yusof, Siti Hajar Leow, Adam Thean Chor Rahman, Raja Noor Zaliha Raja Abd Ngalimat, Mohamad Syazwan Lim, Si Jie Sabri, Suriana |
| author_facet | Yusof, Siti Hajar Leow, Adam Thean Chor Rahman, Raja Noor Zaliha Raja Abd Ngalimat, Mohamad Syazwan Lim, Si Jie Sabri, Suriana |
| author_sort | Yusof, Siti Hajar |
| collection | UPM |
| description | Given its thermoalkaliphilic properties, T1 lipase holds significant potential for diverse industrial applications. However, traditional expression methods in Escherichia coli, specifically the plasmid-based system, present challenges of exerting metabolic burden on host cells and elevated costs due to antibiotic usage. This study addresses these issues by pioneering the expression of recombinant T1 lipase in a sucrose-utilizing E. coli strain W, using molasses as an economical carbon source. The gene cassette (KIKO plasmid), containing the T1 lipase gene regulated by tac and trc promoters, was integrated into the E. coli genome via the λ Red recombinase system. T1 lipase was optimally expressed in shake flasks at 16°C and a 3% molasses concentration in M9 medium with 0.8 mM IPTG as inducer, yielding 0.44 U/mL activity in the genome-based system compared to 0.94 U/mL in the plasmid-based system. This study not only underscores the potential of employing sucrose-utilizing E. coli strain for industrial recombinant protein production but also highlights the need for further optimization of genome-based expression systems. It offers an alternative to reduce costs and enhance sustainability in the stable production of industrially relevant enzymes like T1 lipase, without the need for antibiotic supplementation, and has broader implications for leveraging inexpensive carbon sources like molasses in biotechnological applications. |
| first_indexed | 2024-09-25T03:44:47Z |
| format | Article |
| id | upm.eprints-112021 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-09-25T03:44:47Z |
| publishDate | 2024 |
| publisher | University of Malaya |
| record_format | dspace |
| spelling | upm.eprints-1120212024-09-17T01:26:33Z http://psasir.upm.edu.my/id/eprint/112021/ Plasmid-based and genome-based expression of recombinant T1 lipase in sucrose-utilizing E. coli strain W Yusof, Siti Hajar Leow, Adam Thean Chor Rahman, Raja Noor Zaliha Raja Abd Ngalimat, Mohamad Syazwan Lim, Si Jie Sabri, Suriana Given its thermoalkaliphilic properties, T1 lipase holds significant potential for diverse industrial applications. However, traditional expression methods in Escherichia coli, specifically the plasmid-based system, present challenges of exerting metabolic burden on host cells and elevated costs due to antibiotic usage. This study addresses these issues by pioneering the expression of recombinant T1 lipase in a sucrose-utilizing E. coli strain W, using molasses as an economical carbon source. The gene cassette (KIKO plasmid), containing the T1 lipase gene regulated by tac and trc promoters, was integrated into the E. coli genome via the λ Red recombinase system. T1 lipase was optimally expressed in shake flasks at 16°C and a 3% molasses concentration in M9 medium with 0.8 mM IPTG as inducer, yielding 0.44 U/mL activity in the genome-based system compared to 0.94 U/mL in the plasmid-based system. This study not only underscores the potential of employing sucrose-utilizing E. coli strain for industrial recombinant protein production but also highlights the need for further optimization of genome-based expression systems. It offers an alternative to reduce costs and enhance sustainability in the stable production of industrially relevant enzymes like T1 lipase, without the need for antibiotic supplementation, and has broader implications for leveraging inexpensive carbon sources like molasses in biotechnological applications. University of Malaya 2024-07-30 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/112021/1/112021.pdf Yusof, Siti Hajar and Leow, Adam Thean Chor and Rahman, Raja Noor Zaliha Raja Abd and Ngalimat, Mohamad Syazwan and Lim, Si Jie and Sabri, Suriana (2024) Plasmid-based and genome-based expression of recombinant T1 lipase in sucrose-utilizing E. coli strain W. Asia-Pacific Journal of Molecular Biology and Biotechnology, 32 (3). pp. 113-127. ISSN 0128-7451 https://www.msmbb.my/images/publication/volume_32/issue_3/11-Yusof-et-al.pdf 10.35118/apjmbb.2024.032.3.11 |
| spellingShingle | Yusof, Siti Hajar Leow, Adam Thean Chor Rahman, Raja Noor Zaliha Raja Abd Ngalimat, Mohamad Syazwan Lim, Si Jie Sabri, Suriana Plasmid-based and genome-based expression of recombinant T1 lipase in sucrose-utilizing E. coli strain W |
| title | Plasmid-based and genome-based expression of recombinant T1 lipase in sucrose-utilizing E. coli strain W |
| title_full | Plasmid-based and genome-based expression of recombinant T1 lipase in sucrose-utilizing E. coli strain W |
| title_fullStr | Plasmid-based and genome-based expression of recombinant T1 lipase in sucrose-utilizing E. coli strain W |
| title_full_unstemmed | Plasmid-based and genome-based expression of recombinant T1 lipase in sucrose-utilizing E. coli strain W |
| title_short | Plasmid-based and genome-based expression of recombinant T1 lipase in sucrose-utilizing E. coli strain W |
| title_sort | plasmid based and genome based expression of recombinant t1 lipase in sucrose utilizing e coli strain w |
| url | http://psasir.upm.edu.my/id/eprint/112021/1/112021.pdf |
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