Reductive alkylation of lipase: experimental and molecular modeling approaches
Candida rugosa lipase was modified via reductive alkylation to increase its hydrophobicity to work better in organic solvents. The free amino group of lysines was alkylated using propionaldehyde with different degrees of modification obtained (49 and 86%). Far-ultraviolet circular dichroism (CD) spe...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English English |
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Humana Press
2004
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| Online Access: | http://psasir.upm.edu.my/id/eprint/56252/1/56252.pdf http://psasir.upm.edu.my/id/eprint/56252/7/1-s2.0-014102299190097T-main.pdf |
| _version_ | 1811137490339233792 |
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| author | Raja Abdul Rahman, Raja Noor Zaliha Tejo, Bimo Ario Basri, Mahiran Abdul Rahman, Mohd Basyaruddin Khan, Farid Md Zain, Sharifuddin Siahaan, Teruna J. Salleh, Abu Bakar |
| author_facet | Raja Abdul Rahman, Raja Noor Zaliha Tejo, Bimo Ario Basri, Mahiran Abdul Rahman, Mohd Basyaruddin Khan, Farid Md Zain, Sharifuddin Siahaan, Teruna J. Salleh, Abu Bakar |
| author_sort | Raja Abdul Rahman, Raja Noor Zaliha |
| collection | UPM |
| description | Candida rugosa lipase was modified via reductive alkylation to increase its hydrophobicity to work better in organic solvents. The free amino group of lysines was alkylated using propionaldehyde with different degrees of modification obtained (49 and 86%). Far-ultraviolet circular dichroism (CD) spectroscopy of the lipase in aqueous solvent showed that such chemical modifications at the enzyme surface caused a loss in secondary and tertiary structure that is attributed to the enzyme unfolding. Using molecular modeling, we propose that in an aqueous environment the loss in protein structure of the modified lipase is owing to disruption of stabilizing salt bridges, particularly of surface lysines. Indeed, molecular modeling and simulation of a salt bridge formed by Lys-75 to Asp-79, in a nonpolar environment, suggests the adoption of a more flexible alkylated lysine that may explain higher lipase activity in organic solvents on alkylation. |
| first_indexed | 2024-03-06T09:25:51Z |
| format | Article |
| id | upm.eprints-56252 |
| institution | Universiti Putra Malaysia |
| language | English English |
| last_indexed | 2024-09-25T03:35:07Z |
| publishDate | 2004 |
| publisher | Humana Press |
| record_format | dspace |
| spelling | upm.eprints-562522024-08-05T08:24:37Z http://psasir.upm.edu.my/id/eprint/56252/ Reductive alkylation of lipase: experimental and molecular modeling approaches Raja Abdul Rahman, Raja Noor Zaliha Tejo, Bimo Ario Basri, Mahiran Abdul Rahman, Mohd Basyaruddin Khan, Farid Md Zain, Sharifuddin Siahaan, Teruna J. Salleh, Abu Bakar Candida rugosa lipase was modified via reductive alkylation to increase its hydrophobicity to work better in organic solvents. The free amino group of lysines was alkylated using propionaldehyde with different degrees of modification obtained (49 and 86%). Far-ultraviolet circular dichroism (CD) spectroscopy of the lipase in aqueous solvent showed that such chemical modifications at the enzyme surface caused a loss in secondary and tertiary structure that is attributed to the enzyme unfolding. Using molecular modeling, we propose that in an aqueous environment the loss in protein structure of the modified lipase is owing to disruption of stabilizing salt bridges, particularly of surface lysines. Indeed, molecular modeling and simulation of a salt bridge formed by Lys-75 to Asp-79, in a nonpolar environment, suggests the adoption of a more flexible alkylated lysine that may explain higher lipase activity in organic solvents on alkylation. Humana Press 2004 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/56252/1/56252.pdf text en http://psasir.upm.edu.my/id/eprint/56252/7/1-s2.0-014102299190097T-main.pdf Raja Abdul Rahman, Raja Noor Zaliha and Tejo, Bimo Ario and Basri, Mahiran and Abdul Rahman, Mohd Basyaruddin and Khan, Farid and Md Zain, Sharifuddin and Siahaan, Teruna J. and Salleh, Abu Bakar (2004) Reductive alkylation of lipase: experimental and molecular modeling approaches. Applied Biochemistry and Biotechnology, 118 (1-3). pp. 11-20. ISSN 0273-2289; ESSN: 1559-0291 https://link.springer.com/article/10.1385/ABAB%3A118%3A1-3%3A011 10.1385/ABAB:118:1-3:011 |
| spellingShingle | Raja Abdul Rahman, Raja Noor Zaliha Tejo, Bimo Ario Basri, Mahiran Abdul Rahman, Mohd Basyaruddin Khan, Farid Md Zain, Sharifuddin Siahaan, Teruna J. Salleh, Abu Bakar Reductive alkylation of lipase: experimental and molecular modeling approaches |
| title | Reductive alkylation of lipase: experimental and molecular modeling approaches |
| title_full | Reductive alkylation of lipase: experimental and molecular modeling approaches |
| title_fullStr | Reductive alkylation of lipase: experimental and molecular modeling approaches |
| title_full_unstemmed | Reductive alkylation of lipase: experimental and molecular modeling approaches |
| title_short | Reductive alkylation of lipase: experimental and molecular modeling approaches |
| title_sort | reductive alkylation of lipase experimental and molecular modeling approaches |
| url | http://psasir.upm.edu.my/id/eprint/56252/1/56252.pdf http://psasir.upm.edu.my/id/eprint/56252/7/1-s2.0-014102299190097T-main.pdf |
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