Enhancing the stability of Geobacillus zalihae T1 lipase in organic solvents and insights into the structural stability of its variants
Critical to the applications of proteins in non-aqueous enzymatic processes is their structural dynamics in relation to solvent polarity. A pool of mutants derived from Geobacillus zalihae T1 lipase was screened in organic solvents (methanol, ethanol, propanol, butanol and pentanol) resulting in the...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021
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| Online Access: | http://psasir.upm.edu.my/id/eprint/97110/ http://psasir.upm.edu.my/id/eprint/97110/1/ABSTRACT.pdf |
| _version_ | 1848862517662580736 |
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| author | Maiangwa, Jonathan Hamdan, Siti Hajar Mohamad Ali, Mohd Shukuri Salleh, Abu Bakar Raja Abd Rahman, Raja Noor Zaliha Mohd Shariff, Fairolniza Leow, Adam Thean Chor |
| author_facet | Maiangwa, Jonathan Hamdan, Siti Hajar Mohamad Ali, Mohd Shukuri Salleh, Abu Bakar Raja Abd Rahman, Raja Noor Zaliha Mohd Shariff, Fairolniza Leow, Adam Thean Chor |
| author_sort | Maiangwa, Jonathan |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Critical to the applications of proteins in non-aqueous enzymatic processes is their structural dynamics in relation to solvent polarity. A pool of mutants derived from Geobacillus zalihae T1 lipase was screened in organic solvents (methanol, ethanol, propanol, butanol and pentanol) resulting in the selection of six mutants at initial screening (A83D/K251E, R21C, G35D/S195 N, K84R/R103C/M121I/T272 M and R106H/G327S). Site-directed mutagenesis further yielded quadruple mutants A83D/M121I/K251E/G327S and A83D/M121I/S195 N/T272 M, both of which had improved activity after incubation in methanol. The km and kcat values of these mutants vary marginally with the wild-type enzyme in the methanol/substrate mixture. Thermally induced unfolding of mutants was accompanied with some loss of secondary structure content. The root mean square deviations (RMSD) and B-factors revealed that changes in the structural organization are intertwined with an interplay of the protein backbone with organic solvents. Spatially exposed charged residues showed correlations between the solvation dynamics of the methanol solvent and the hydrophobicity of the residues. The short distances of the radial distribution function provided the required distances for hydrogen bond formation and hydrophobic interactions. These dynamic changes demonstrate newly formed structural interactions could be targeted and incorporated experimentally on the basis of solvent mobility and mutant residues. |
| first_indexed | 2025-11-15T13:18:17Z |
| format | Article |
| id | upm-97110 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T13:18:17Z |
| publishDate | 2021 |
| publisher | Elsevier |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-971102022-10-15T14:44:54Z http://psasir.upm.edu.my/id/eprint/97110/ Enhancing the stability of Geobacillus zalihae T1 lipase in organic solvents and insights into the structural stability of its variants Maiangwa, Jonathan Hamdan, Siti Hajar Mohamad Ali, Mohd Shukuri Salleh, Abu Bakar Raja Abd Rahman, Raja Noor Zaliha Mohd Shariff, Fairolniza Leow, Adam Thean Chor Critical to the applications of proteins in non-aqueous enzymatic processes is their structural dynamics in relation to solvent polarity. A pool of mutants derived from Geobacillus zalihae T1 lipase was screened in organic solvents (methanol, ethanol, propanol, butanol and pentanol) resulting in the selection of six mutants at initial screening (A83D/K251E, R21C, G35D/S195 N, K84R/R103C/M121I/T272 M and R106H/G327S). Site-directed mutagenesis further yielded quadruple mutants A83D/M121I/K251E/G327S and A83D/M121I/S195 N/T272 M, both of which had improved activity after incubation in methanol. The km and kcat values of these mutants vary marginally with the wild-type enzyme in the methanol/substrate mixture. Thermally induced unfolding of mutants was accompanied with some loss of secondary structure content. The root mean square deviations (RMSD) and B-factors revealed that changes in the structural organization are intertwined with an interplay of the protein backbone with organic solvents. Spatially exposed charged residues showed correlations between the solvation dynamics of the methanol solvent and the hydrophobicity of the residues. The short distances of the radial distribution function provided the required distances for hydrogen bond formation and hydrophobic interactions. These dynamic changes demonstrate newly formed structural interactions could be targeted and incorporated experimentally on the basis of solvent mobility and mutant residues. Elsevier 2021 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/97110/1/ABSTRACT.pdf Maiangwa, Jonathan and Hamdan, Siti Hajar and Mohamad Ali, Mohd Shukuri and Salleh, Abu Bakar and Raja Abd Rahman, Raja Noor Zaliha and Mohd Shariff, Fairolniza and Leow, Adam Thean Chor (2021) Enhancing the stability of Geobacillus zalihae T1 lipase in organic solvents and insights into the structural stability of its variants. Journal of Molecular Graphics and Modelling, 105. art. no. 107897. pp. 1-14. ISSN 1093-3263; ESSN: 1873-4243 https://www.sciencedirect.com/science/article/abs/pii/S1093326321000668 10.1016/j.jmgm.2021.107897 |
| spellingShingle | Maiangwa, Jonathan Hamdan, Siti Hajar Mohamad Ali, Mohd Shukuri Salleh, Abu Bakar Raja Abd Rahman, Raja Noor Zaliha Mohd Shariff, Fairolniza Leow, Adam Thean Chor Enhancing the stability of Geobacillus zalihae T1 lipase in organic solvents and insights into the structural stability of its variants |
| title | Enhancing the stability of Geobacillus zalihae T1 lipase in organic solvents and insights into the structural stability of its variants |
| title_full | Enhancing the stability of Geobacillus zalihae T1 lipase in organic solvents and insights into the structural stability of its variants |
| title_fullStr | Enhancing the stability of Geobacillus zalihae T1 lipase in organic solvents and insights into the structural stability of its variants |
| title_full_unstemmed | Enhancing the stability of Geobacillus zalihae T1 lipase in organic solvents and insights into the structural stability of its variants |
| title_short | Enhancing the stability of Geobacillus zalihae T1 lipase in organic solvents and insights into the structural stability of its variants |
| title_sort | enhancing the stability of geobacillus zalihae t1 lipase in organic solvents and insights into the structural stability of its variants |
| url | http://psasir.upm.edu.my/id/eprint/97110/ http://psasir.upm.edu.my/id/eprint/97110/ http://psasir.upm.edu.my/id/eprint/97110/ http://psasir.upm.edu.my/id/eprint/97110/1/ABSTRACT.pdf |