Single residue substitution at N-terminal affects temperature stability and activity of L2 lipase
Rational design is widely employed in protein engineering to tailor wild-type enzymes for industrial applications. The typical target region for mutation is a functional region like the catalytic site to improve stability and activity. However, few have explored the role of other regions which, in p...
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| Format: | Article |
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MDPI AG
2020
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| author | Bukhari, Noramirah Leow, Adam Thean Chor Raja Abdul Rahman, Raja Noor Zaliha Mohd Shariff, Fairolniza |
| author_facet | Bukhari, Noramirah Leow, Adam Thean Chor Raja Abdul Rahman, Raja Noor Zaliha Mohd Shariff, Fairolniza |
| author_sort | Bukhari, Noramirah |
| collection | UPM |
| description | Rational design is widely employed in protein engineering to tailor wild-type enzymes for industrial applications. The typical target region for mutation is a functional region like the catalytic site to improve stability and activity. However, few have explored the role of other regions which, in principle, have no evident functionality such as the N-terminal region. In this study, stability prediction software was used to identify the critical point in the non-functional N-terminal region of L2 lipase and the effects of the substitution towards temperature stability and activity were determined. The results showed 3 mutant lipases: A8V, A8P and A8E with 29% better thermostability, 4 h increase in half-life and 6.6 °C higher thermal denaturation point, respectively. A8V showed 1.6-fold enhancement in activity compared to wild-type. To conclude, the improvement in temperature stability upon substitution showed that the N-terminal region plays a role in temperature stability and activity of L2 lipase. |
| first_indexed | 2024-03-06T10:41:21Z |
| format | Article |
| id | upm.eprints-85827 |
| institution | Universiti Putra Malaysia |
| last_indexed | 2024-03-06T10:41:21Z |
| publishDate | 2020 |
| publisher | MDPI AG |
| record_format | dspace |
| spelling | upm.eprints-858272023-10-02T03:56:34Z http://psasir.upm.edu.my/id/eprint/85827/ Single residue substitution at N-terminal affects temperature stability and activity of L2 lipase Bukhari, Noramirah Leow, Adam Thean Chor Raja Abdul Rahman, Raja Noor Zaliha Mohd Shariff, Fairolniza Rational design is widely employed in protein engineering to tailor wild-type enzymes for industrial applications. The typical target region for mutation is a functional region like the catalytic site to improve stability and activity. However, few have explored the role of other regions which, in principle, have no evident functionality such as the N-terminal region. In this study, stability prediction software was used to identify the critical point in the non-functional N-terminal region of L2 lipase and the effects of the substitution towards temperature stability and activity were determined. The results showed 3 mutant lipases: A8V, A8P and A8E with 29% better thermostability, 4 h increase in half-life and 6.6 °C higher thermal denaturation point, respectively. A8V showed 1.6-fold enhancement in activity compared to wild-type. To conclude, the improvement in temperature stability upon substitution showed that the N-terminal region plays a role in temperature stability and activity of L2 lipase. MDPI AG 2020 Article PeerReviewed Bukhari, Noramirah and Leow, Adam Thean Chor and Raja Abdul Rahman, Raja Noor Zaliha and Mohd Shariff, Fairolniza (2020) Single residue substitution at N-terminal affects temperature stability and activity of L2 lipase. Molecules, 25 (15). art. no. 3433. pp. 1-18. ISSN 1420-3049 https://www.mdpi.com/1420-3049/25/15/3433 10.3390/molecules25153433 |
| spellingShingle | Bukhari, Noramirah Leow, Adam Thean Chor Raja Abdul Rahman, Raja Noor Zaliha Mohd Shariff, Fairolniza Single residue substitution at N-terminal affects temperature stability and activity of L2 lipase |
| title | Single residue substitution at N-terminal affects temperature stability and activity of L2 lipase |
| title_full | Single residue substitution at N-terminal affects temperature stability and activity of L2 lipase |
| title_fullStr | Single residue substitution at N-terminal affects temperature stability and activity of L2 lipase |
| title_full_unstemmed | Single residue substitution at N-terminal affects temperature stability and activity of L2 lipase |
| title_short | Single residue substitution at N-terminal affects temperature stability and activity of L2 lipase |
| title_sort | single residue substitution at n terminal affects temperature stability and activity of l2 lipase |
| work_keys_str_mv | AT bukharinoramirah singleresiduesubstitutionatnterminalaffectstemperaturestabilityandactivityofl2lipase AT leowadamtheanchor singleresiduesubstitutionatnterminalaffectstemperaturestabilityandactivityofl2lipase AT rajaabdulrahmanrajanoorzaliha singleresiduesubstitutionatnterminalaffectstemperaturestabilityandactivityofl2lipase AT mohdsharifffairolniza singleresiduesubstitutionatnterminalaffectstemperaturestabilityandactivityofl2lipase |