Facile modulation of enantioselectivity of thermophilic Geobacillus zalihae lipase by regulating hydrophobicity of its Q114 oxyanion
Site-directed mutagenesis of the oxyanion-containing amino acid Q114 in the recombinant thermophilic T1 lipase previously isolated from Geobacillus zalihae was performed to elucidate its role in the enzyme⿿s enantioselectivity and reactivity. Substitution of Q114 with a hydrophobic methionine to yie...
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| Format: | Article |
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Elsevier
2016
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| Online Access: | http://psasir.upm.edu.my/id/eprint/54264/ http://psasir.upm.edu.my/id/eprint/54264/1/Facile%20modulation%20of%20enantioselectivity%20of%20thermophilic%20Geobacillus%20zalihae%20lipase%20by%20regulating%20hydrophobicity%20of%20its%20Q114%20oxyanion.pdf |
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| author | Abdul Wahab, Roswanira Basri, Mahiran Raja Abdul Rahman, Raja Noor Zaliha Salleh, Abu Bakar Abdul Rahman, Mohd Basyaruddin Leow, Adam Thean Chor |
| author_facet | Abdul Wahab, Roswanira Basri, Mahiran Raja Abdul Rahman, Raja Noor Zaliha Salleh, Abu Bakar Abdul Rahman, Mohd Basyaruddin Leow, Adam Thean Chor |
| author_sort | Abdul Wahab, Roswanira |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Site-directed mutagenesis of the oxyanion-containing amino acid Q114 in the recombinant thermophilic T1 lipase previously isolated from Geobacillus zalihae was performed to elucidate its role in the enzyme⿿s enantioselectivity and reactivity. Substitution of Q114 with a hydrophobic methionine to yield mutant Q114M increased enantioselectivity (3.2-fold) and marginally improved reactivity (1.4-fold) of the lipase in catalysing esterification of ibuprofen with oleyl alcohol. The improved catalytic efficiency of Q114L was concomitant with reduced flexibility in the active site while the decreased enantioselectivity of Q114L could be directly attributed to diminished electrostatic repulsion of the substrate carboxylate ion that rendered partial loss in steric hindrance and thus enantioselectivity. The highest E-values for both Q114L (E-value 14.6) and Q114M (E-value 48.5) mutant lipases were attained at 50 °C, after 12⿿16 h, with a molar ratio of oleyl alcohol to ibuprofen of 1.5:1 and at 2.0% (w/v) enzyme load without addition of molecular sieves. Pertinently, site-directed mutagenesis on the Q114 oxyanion of T1 resulted in improved enantioselectivity and such approach may be applicable to other lipases of the same family. We demonstrated that electrostatic repulsion phenomena could affect flexibility/rigidity of the enzyme-substrate complex, aspects vital for enzyme activity and enantioselectivity of T1. |
| first_indexed | 2025-11-15T10:39:01Z |
| format | Article |
| id | upm-54264 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T10:39:01Z |
| publishDate | 2016 |
| publisher | Elsevier |
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| spelling | upm-542642018-03-09T03:09:58Z http://psasir.upm.edu.my/id/eprint/54264/ Facile modulation of enantioselectivity of thermophilic Geobacillus zalihae lipase by regulating hydrophobicity of its Q114 oxyanion Abdul Wahab, Roswanira Basri, Mahiran Raja Abdul Rahman, Raja Noor Zaliha Salleh, Abu Bakar Abdul Rahman, Mohd Basyaruddin Leow, Adam Thean Chor Site-directed mutagenesis of the oxyanion-containing amino acid Q114 in the recombinant thermophilic T1 lipase previously isolated from Geobacillus zalihae was performed to elucidate its role in the enzyme⿿s enantioselectivity and reactivity. Substitution of Q114 with a hydrophobic methionine to yield mutant Q114M increased enantioselectivity (3.2-fold) and marginally improved reactivity (1.4-fold) of the lipase in catalysing esterification of ibuprofen with oleyl alcohol. The improved catalytic efficiency of Q114L was concomitant with reduced flexibility in the active site while the decreased enantioselectivity of Q114L could be directly attributed to diminished electrostatic repulsion of the substrate carboxylate ion that rendered partial loss in steric hindrance and thus enantioselectivity. The highest E-values for both Q114L (E-value 14.6) and Q114M (E-value 48.5) mutant lipases were attained at 50 °C, after 12⿿16 h, with a molar ratio of oleyl alcohol to ibuprofen of 1.5:1 and at 2.0% (w/v) enzyme load without addition of molecular sieves. Pertinently, site-directed mutagenesis on the Q114 oxyanion of T1 resulted in improved enantioselectivity and such approach may be applicable to other lipases of the same family. We demonstrated that electrostatic repulsion phenomena could affect flexibility/rigidity of the enzyme-substrate complex, aspects vital for enzyme activity and enantioselectivity of T1. Elsevier 2016-11 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/54264/1/Facile%20modulation%20of%20enantioselectivity%20of%20thermophilic%20Geobacillus%20zalihae%20lipase%20by%20regulating%20hydrophobicity%20of%20its%20Q114%20oxyanion.pdf Abdul Wahab, Roswanira and Basri, Mahiran and Raja Abdul Rahman, Raja Noor Zaliha and Salleh, Abu Bakar and Abdul Rahman, Mohd Basyaruddin and Leow, Adam Thean Chor (2016) Facile modulation of enantioselectivity of thermophilic Geobacillus zalihae lipase by regulating hydrophobicity of its Q114 oxyanion. Enzyme and Microbial Technology, 93-94. pp. 174-181. ISSN 0141-0229; ESSN: 1879-0909 https://www.sciencedirect.com/science/article/pii/S0141022916301715 T1 lipase; Geobacillus zalihae; 2DSN; Site-directed mutagenesis; Enantioselectivity; Oxyanion 10.1016/j.enzmictec.2016.08.020 |
| spellingShingle | T1 lipase; Geobacillus zalihae; 2DSN; Site-directed mutagenesis; Enantioselectivity; Oxyanion Abdul Wahab, Roswanira Basri, Mahiran Raja Abdul Rahman, Raja Noor Zaliha Salleh, Abu Bakar Abdul Rahman, Mohd Basyaruddin Leow, Adam Thean Chor Facile modulation of enantioselectivity of thermophilic Geobacillus zalihae lipase by regulating hydrophobicity of its Q114 oxyanion |
| title | Facile modulation of enantioselectivity of thermophilic Geobacillus zalihae lipase by regulating hydrophobicity of its Q114 oxyanion |
| title_full | Facile modulation of enantioselectivity of thermophilic Geobacillus zalihae lipase by regulating hydrophobicity of its Q114 oxyanion |
| title_fullStr | Facile modulation of enantioselectivity of thermophilic Geobacillus zalihae lipase by regulating hydrophobicity of its Q114 oxyanion |
| title_full_unstemmed | Facile modulation of enantioselectivity of thermophilic Geobacillus zalihae lipase by regulating hydrophobicity of its Q114 oxyanion |
| title_short | Facile modulation of enantioselectivity of thermophilic Geobacillus zalihae lipase by regulating hydrophobicity of its Q114 oxyanion |
| title_sort | facile modulation of enantioselectivity of thermophilic geobacillus zalihae lipase by regulating hydrophobicity of its q114 oxyanion |
| topic | T1 lipase; Geobacillus zalihae; 2DSN; Site-directed mutagenesis; Enantioselectivity; Oxyanion |
| url | http://psasir.upm.edu.my/id/eprint/54264/ http://psasir.upm.edu.my/id/eprint/54264/ http://psasir.upm.edu.my/id/eprint/54264/ http://psasir.upm.edu.my/id/eprint/54264/1/Facile%20modulation%20of%20enantioselectivity%20of%20thermophilic%20Geobacillus%20zalihae%20lipase%20by%20regulating%20hydrophobicity%20of%20its%20Q114%20oxyanion.pdf |