Reductive alkylation causes the formation of a molten globule-like intermediate structure in Geobacillus zalihae strain T1 thermostable lipase
A thermostable lipase from Geobacillus zalihae strain T1 was chemically modified using propionaldehyde via reductive alkylation. The targeted alkylation sites were lysines, in which T1 lipase possessed 11 residues. Far-UV circular dichroism (CD) spectra of both native and alkylated enzyme showed a s...
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| Format: | Article |
| Language: | English |
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Springer
2011
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| Online Access: | http://psasir.upm.edu.my/id/eprint/22284/1/Reductive%20alkylation%20causes%20the%20formation%20of%20a%20molten%20globule.pdf |
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| author | Cheong, Kok Whye Leow, Adam Thean Chor Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Abdul Rahman, Mohd Basyaruddin Salleh, Abu Bakar |
| author_facet | Cheong, Kok Whye Leow, Adam Thean Chor Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Abdul Rahman, Mohd Basyaruddin Salleh, Abu Bakar |
| author_sort | Cheong, Kok Whye |
| collection | UPM |
| description | A thermostable lipase from Geobacillus zalihae strain T1 was chemically modified using propionaldehyde via reductive alkylation. The targeted alkylation sites were lysines, in which T1 lipase possessed 11 residues. Far-UV circular dichroism (CD) spectra of both native and alkylated enzyme showed a similar broad minimum between 208 and 222 nm, thus suggesting a substantial amount of secondary structures in modified enzyme, as compared with the corresponding native enzyme. The hydrolytic activity of the modified enzymes dropped drastically by nearly 15-fold upon chemical modification, despite both the native and modified form showed distinctive α-helical bands at 208 and 222 nm in CD spectra, leading us to the hypothesis of formation of a molten globule (MG)-like structure. As cooperative unfolding transitions were observed, the modified lipase was distinguished from the native state, in which the former possessed a denaturation temperature (T m) in lower temperature range at 61 °C while the latter at 68 °C. This was further supported by 8-anilino-1-naphthalenesulfonic acid (ANS) probed fluorescence which indicated higher exposure of hydrophobic residues, consequential of chemical modification. Based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis, a small number of lysine residues were confirmed to be alkylated. |
| first_indexed | 2024-03-06T07:53:30Z |
| format | Article |
| id | upm.eprints-22284 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T07:53:30Z |
| publishDate | 2011 |
| publisher | Springer |
| record_format | dspace |
| spelling | upm.eprints-222842016-09-27T08:54:42Z http://psasir.upm.edu.my/id/eprint/22284/ Reductive alkylation causes the formation of a molten globule-like intermediate structure in Geobacillus zalihae strain T1 thermostable lipase Cheong, Kok Whye Leow, Adam Thean Chor Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Abdul Rahman, Mohd Basyaruddin Salleh, Abu Bakar A thermostable lipase from Geobacillus zalihae strain T1 was chemically modified using propionaldehyde via reductive alkylation. The targeted alkylation sites were lysines, in which T1 lipase possessed 11 residues. Far-UV circular dichroism (CD) spectra of both native and alkylated enzyme showed a similar broad minimum between 208 and 222 nm, thus suggesting a substantial amount of secondary structures in modified enzyme, as compared with the corresponding native enzyme. The hydrolytic activity of the modified enzymes dropped drastically by nearly 15-fold upon chemical modification, despite both the native and modified form showed distinctive α-helical bands at 208 and 222 nm in CD spectra, leading us to the hypothesis of formation of a molten globule (MG)-like structure. As cooperative unfolding transitions were observed, the modified lipase was distinguished from the native state, in which the former possessed a denaturation temperature (T m) in lower temperature range at 61 °C while the latter at 68 °C. This was further supported by 8-anilino-1-naphthalenesulfonic acid (ANS) probed fluorescence which indicated higher exposure of hydrophobic residues, consequential of chemical modification. Based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis, a small number of lysine residues were confirmed to be alkylated. Springer 2011-06 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/22284/1/Reductive%20alkylation%20causes%20the%20formation%20of%20a%20molten%20globule.pdf Cheong, Kok Whye and Leow, Adam Thean Chor and Raja Abdul Rahman, Raja Noor Zaliha and Basri, Mahiran and Abdul Rahman, Mohd Basyaruddin and Salleh, Abu Bakar (2011) Reductive alkylation causes the formation of a molten globule-like intermediate structure in Geobacillus zalihae strain T1 thermostable lipase. Applied Biochemistry and Biotechnology, 164 (3). pp. 362-375. ISSN 0273-2289; ESSN:1559-0291 http://link.springer.com/article/10.1007/s12010-010-9140-8?view=classic 10.1007/s12010-010-9140-8 |
| spellingShingle | Cheong, Kok Whye Leow, Adam Thean Chor Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Abdul Rahman, Mohd Basyaruddin Salleh, Abu Bakar Reductive alkylation causes the formation of a molten globule-like intermediate structure in Geobacillus zalihae strain T1 thermostable lipase |
| title | Reductive alkylation causes the formation of a molten globule-like intermediate structure in Geobacillus zalihae strain T1 thermostable lipase |
| title_full | Reductive alkylation causes the formation of a molten globule-like intermediate structure in Geobacillus zalihae strain T1 thermostable lipase |
| title_fullStr | Reductive alkylation causes the formation of a molten globule-like intermediate structure in Geobacillus zalihae strain T1 thermostable lipase |
| title_full_unstemmed | Reductive alkylation causes the formation of a molten globule-like intermediate structure in Geobacillus zalihae strain T1 thermostable lipase |
| title_short | Reductive alkylation causes the formation of a molten globule-like intermediate structure in Geobacillus zalihae strain T1 thermostable lipase |
| title_sort | reductive alkylation causes the formation of a molten globule like intermediate structure in geobacillus zalihae strain t1 thermostable lipase |
| url | http://psasir.upm.edu.my/id/eprint/22284/1/Reductive%20alkylation%20causes%20the%20formation%20of%20a%20molten%20globule.pdf |
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