Exploring the impacts of terminal mutations on the thermostability of Bacillus sp. L2 lipase
Protein engineering has been widely used to improve enzyme properties and make them appropriate for use as industrial biocatalysts. To study the effect of mutation at the N-and C-terminal of lipase, two double mutants (A8V/S385E and A8P/S385E) were generated by site-directed mutagenesis of L2 lipase...
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| Format: | Article |
| Language: | English |
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University of Malaya
2025
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| Online Access: | http://psasir.upm.edu.my/id/eprint/121194/ http://psasir.upm.edu.my/id/eprint/121194/1/121194.pdf |
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| author | Ishak, Siti Nor Hasmah Mohd Faizal, Khairina Alya Anasir, Mohd Ishtiaq Bukhari, Noramirah Zulkifli Lubis, Nur Aisyah Mat Radzi, Mohd Radzman Leow, Adam Thean Chor Mohamad Ali, Mohd Shukuri Mohd Shariff, Fairolniza |
| author_facet | Ishak, Siti Nor Hasmah Mohd Faizal, Khairina Alya Anasir, Mohd Ishtiaq Bukhari, Noramirah Zulkifli Lubis, Nur Aisyah Mat Radzi, Mohd Radzman Leow, Adam Thean Chor Mohamad Ali, Mohd Shukuri Mohd Shariff, Fairolniza |
| author_sort | Ishak, Siti Nor Hasmah |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Protein engineering has been widely used to improve enzyme properties and make them appropriate for use as industrial biocatalysts. To study the effect of mutation at the N-and C-terminal of lipase, two double mutants (A8V/S385E and A8P/S385E) were generated by site-directed mutagenesis of L2 lipase from Bacillus sp. (wt-L2). The simultaneous mutations in the A8V/S385E and A8P/S385E resulted in significant changes in the lipase's properties compared to the wild-type (wt-L2). The mutants demonstrated increased thermostability compared to the wild-type. The melting temperature (Tm) analysis using circular dichroism revealed higher Tm values of 84.5 °C for A8P/S385E and 75.1 °C for A8V/S385E. This indicates that the enzyme can withstand higher temperatures before denaturation, a desirable trait in various industrial processes. Secondary structure analysis indicated alterations in the lipase structure caused by the simultaneous mutations. In summary, the simultaneous mutation at the C-and N-terminals had a multifaceted impact on the lipase, influencing its optimal temperature, thermostability, and structural characteristics. These findings provide insights into how specific genetic modifications can be employed to tailor the enzyme for improved performance in industrial applications. |
| first_indexed | 2025-11-15T14:50:29Z |
| format | Article |
| id | upm-121194 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T14:50:29Z |
| publishDate | 2025 |
| publisher | University of Malaya |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-1211942025-10-29T01:13:56Z http://psasir.upm.edu.my/id/eprint/121194/ Exploring the impacts of terminal mutations on the thermostability of Bacillus sp. L2 lipase Ishak, Siti Nor Hasmah Mohd Faizal, Khairina Alya Anasir, Mohd Ishtiaq Bukhari, Noramirah Zulkifli Lubis, Nur Aisyah Mat Radzi, Mohd Radzman Leow, Adam Thean Chor Mohamad Ali, Mohd Shukuri Mohd Shariff, Fairolniza Protein engineering has been widely used to improve enzyme properties and make them appropriate for use as industrial biocatalysts. To study the effect of mutation at the N-and C-terminal of lipase, two double mutants (A8V/S385E and A8P/S385E) were generated by site-directed mutagenesis of L2 lipase from Bacillus sp. (wt-L2). The simultaneous mutations in the A8V/S385E and A8P/S385E resulted in significant changes in the lipase's properties compared to the wild-type (wt-L2). The mutants demonstrated increased thermostability compared to the wild-type. The melting temperature (Tm) analysis using circular dichroism revealed higher Tm values of 84.5 °C for A8P/S385E and 75.1 °C for A8V/S385E. This indicates that the enzyme can withstand higher temperatures before denaturation, a desirable trait in various industrial processes. Secondary structure analysis indicated alterations in the lipase structure caused by the simultaneous mutations. In summary, the simultaneous mutation at the C-and N-terminals had a multifaceted impact on the lipase, influencing its optimal temperature, thermostability, and structural characteristics. These findings provide insights into how specific genetic modifications can be employed to tailor the enzyme for improved performance in industrial applications. University of Malaya 2025-06-30 Article PeerReviewed text en cc_by_nc_4 http://psasir.upm.edu.my/id/eprint/121194/1/121194.pdf Ishak, Siti Nor Hasmah and Mohd Faizal, Khairina Alya and Anasir, Mohd Ishtiaq and Bukhari, Noramirah and Zulkifli Lubis, Nur Aisyah and Mat Radzi, Mohd Radzman and Leow, Adam Thean Chor and Mohamad Ali, Mohd Shukuri and Mohd Shariff, Fairolniza (2025) Exploring the impacts of terminal mutations on the thermostability of Bacillus sp. L2 lipase. Asia-Pacific Journal of Molecular Biology and Biotechnology, 33 (2). pp. 144-153. ISSN 0128-7451; eISSN: 2672-7277 https://www.msmbb.my/images/publication/volume_33/issue_2/15-Ishak-et-al.pdf 10.35118/apjmbb.2025.033.2.15 |
| spellingShingle | Ishak, Siti Nor Hasmah Mohd Faizal, Khairina Alya Anasir, Mohd Ishtiaq Bukhari, Noramirah Zulkifli Lubis, Nur Aisyah Mat Radzi, Mohd Radzman Leow, Adam Thean Chor Mohamad Ali, Mohd Shukuri Mohd Shariff, Fairolniza Exploring the impacts of terminal mutations on the thermostability of Bacillus sp. L2 lipase |
| title | Exploring the impacts of terminal mutations on the thermostability of Bacillus sp. L2 lipase |
| title_full | Exploring the impacts of terminal mutations on the thermostability of Bacillus sp. L2 lipase |
| title_fullStr | Exploring the impacts of terminal mutations on the thermostability of Bacillus sp. L2 lipase |
| title_full_unstemmed | Exploring the impacts of terminal mutations on the thermostability of Bacillus sp. L2 lipase |
| title_short | Exploring the impacts of terminal mutations on the thermostability of Bacillus sp. L2 lipase |
| title_sort | exploring the impacts of terminal mutations on the thermostability of bacillus sp. l2 lipase |
| url | http://psasir.upm.edu.my/id/eprint/121194/ http://psasir.upm.edu.my/id/eprint/121194/ http://psasir.upm.edu.my/id/eprint/121194/ http://psasir.upm.edu.my/id/eprint/121194/1/121194.pdf |