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Combination of Oxyanion Gln114 Mutation and Medium Engineering to Influence the Enantioselectivity of Thermophilic Lipase from Geobacillus zalihae

The substitution of the oxyanion Q114 with Met and Leu was carried out to investigate the role of Q114 in imparting enantioselectivity on T1 lipase. The mutation improved enantioselectivity in Q114M over the wild-type, while enantioselectivity in Q114L was reduced. The enantioselectivity of the ther...

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Autores principales: Wahab, Roswanira Abdul, Basri, Mahiran, Rahman, Mohd Basyaruddin Abdul, Rahman, Raja Noor Zaliha Raja Abdul, Salleh, Abu Bakar, Leow, Thean Chor
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3472768/
https://www.ncbi.nlm.nih.gov/pubmed/23109876
http://dx.doi.org/10.3390/ijms130911666
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author Wahab, Roswanira Abdul
Basri, Mahiran
Rahman, Mohd Basyaruddin Abdul
Rahman, Raja Noor Zaliha Raja Abdul
Salleh, Abu Bakar
Leow, Thean Chor
author_facet Wahab, Roswanira Abdul
Basri, Mahiran
Rahman, Mohd Basyaruddin Abdul
Rahman, Raja Noor Zaliha Raja Abdul
Salleh, Abu Bakar
Leow, Thean Chor
author_sort Wahab, Roswanira Abdul
collection PubMed
description The substitution of the oxyanion Q114 with Met and Leu was carried out to investigate the role of Q114 in imparting enantioselectivity on T1 lipase. The mutation improved enantioselectivity in Q114M over the wild-type, while enantioselectivity in Q114L was reduced. The enantioselectivity of the thermophilic lipases, T1, Q114L and Q114M correlated better with log p as compared to the dielectric constant and dipole moment of the solvents. Enzyme activity was good in solvents with log p < 3.5, with the exception of hexane which deviated substantially. Isooctane was found to be the best solvent for the esterification of (R,S)-ibuprofen with oleyl alcohol for lipases Q114M and Q114L, to afford E values of 53.7 and 12.2, respectively. Selectivity of T1 was highest in tetradecane with E value 49.2. Solvents with low log p reduced overall lipase activity and dimethyl sulfoxide (DMSO) completely inhibited the lipases. Ester conversions, however, were still low. Molecular sieves employed as desiccant were found to adversely affect catalysis in the lipase variants, particularly in Q114M. The higher desiccant loading also increased viscosity in the reaction and further reduced the efficiency of the lipase-catalyzed esterifications.
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spelling pubmed-34727682012-10-29 Combination of Oxyanion Gln114 Mutation and Medium Engineering to Influence the Enantioselectivity of Thermophilic Lipase from Geobacillus zalihae Wahab, Roswanira Abdul Basri, Mahiran Rahman, Mohd Basyaruddin Abdul Rahman, Raja Noor Zaliha Raja Abdul Salleh, Abu Bakar Leow, Thean Chor Int J Mol Sci Article The substitution of the oxyanion Q114 with Met and Leu was carried out to investigate the role of Q114 in imparting enantioselectivity on T1 lipase. The mutation improved enantioselectivity in Q114M over the wild-type, while enantioselectivity in Q114L was reduced. The enantioselectivity of the thermophilic lipases, T1, Q114L and Q114M correlated better with log p as compared to the dielectric constant and dipole moment of the solvents. Enzyme activity was good in solvents with log p < 3.5, with the exception of hexane which deviated substantially. Isooctane was found to be the best solvent for the esterification of (R,S)-ibuprofen with oleyl alcohol for lipases Q114M and Q114L, to afford E values of 53.7 and 12.2, respectively. Selectivity of T1 was highest in tetradecane with E value 49.2. Solvents with low log p reduced overall lipase activity and dimethyl sulfoxide (DMSO) completely inhibited the lipases. Ester conversions, however, were still low. Molecular sieves employed as desiccant were found to adversely affect catalysis in the lipase variants, particularly in Q114M. The higher desiccant loading also increased viscosity in the reaction and further reduced the efficiency of the lipase-catalyzed esterifications. Molecular Diversity Preservation International (MDPI) 2012-09-17 /pmc/articles/PMC3472768/ /pubmed/23109876 http://dx.doi.org/10.3390/ijms130911666 Text en © 2012 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0 This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Wahab, Roswanira Abdul
Basri, Mahiran
Rahman, Mohd Basyaruddin Abdul
Rahman, Raja Noor Zaliha Raja Abdul
Salleh, Abu Bakar
Leow, Thean Chor
Combination of Oxyanion Gln114 Mutation and Medium Engineering to Influence the Enantioselectivity of Thermophilic Lipase from Geobacillus zalihae
title Combination of Oxyanion Gln114 Mutation and Medium Engineering to Influence the Enantioselectivity of Thermophilic Lipase from Geobacillus zalihae
title_full Combination of Oxyanion Gln114 Mutation and Medium Engineering to Influence the Enantioselectivity of Thermophilic Lipase from Geobacillus zalihae
title_fullStr Combination of Oxyanion Gln114 Mutation and Medium Engineering to Influence the Enantioselectivity of Thermophilic Lipase from Geobacillus zalihae
title_full_unstemmed Combination of Oxyanion Gln114 Mutation and Medium Engineering to Influence the Enantioselectivity of Thermophilic Lipase from Geobacillus zalihae
title_short Combination of Oxyanion Gln114 Mutation and Medium Engineering to Influence the Enantioselectivity of Thermophilic Lipase from Geobacillus zalihae
title_sort combination of oxyanion gln114 mutation and medium engineering to influence the enantioselectivity of thermophilic lipase from geobacillus zalihae
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3472768/
https://www.ncbi.nlm.nih.gov/pubmed/23109876
http://dx.doi.org/10.3390/ijms130911666
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