The Effect of N-Terminal Domain Removal towards the Biochemical and Structural Features of a Thermotolerant Lipase from an Antarctic Pseudomonas sp. Strain AMS3
Lipase plays an important role in industrial and biotechnological applications. Lipases have been subject to modification at the N and C terminals, allowing better understanding of lipase stability and the discovery of novel properties. A thermotolerant lipase has been isolated from Antarctic Pseudo...
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ftmdpi:oai:mdpi.com:/1422-0067/19/2/560/ 2023-08-20T04:01:39+02:00 The Effect of N-Terminal Domain Removal towards the Biochemical and Structural Features of a Thermotolerant Lipase from an Antarctic Pseudomonas sp. Strain AMS3 Wahhida Latip Raja Raja Abd Rahman Adam Leow Fairolniza Mohd Shariff Nor Kamarudin Mohd Mohamad Ali agris 2018-02-13 application/pdf https://doi.org/10.3390/ijms19020560 EN eng Multidisciplinary Digital Publishing Institute Biochemistry https://dx.doi.org/10.3390/ijms19020560 https://creativecommons.org/licenses/by/4.0/ International Journal of Molecular Sciences; Volume 19; Issue 2; Pages: 560 microbial enzyme N-terminal domain biochemical characterization in silico lipase antarctic Text 2018 ftmdpi https://doi.org/10.3390/ijms19020560 2023-07-31T21:23:26Z Lipase plays an important role in industrial and biotechnological applications. Lipases have been subject to modification at the N and C terminals, allowing better understanding of lipase stability and the discovery of novel properties. A thermotolerant lipase has been isolated from Antarctic Pseudomonas sp. The purified Antarctic AMS3 lipase (native) was found to be stable across a broad range of temperatures and pH levels. The lipase has a partial Glutathione-S-transferase type C (GST-C) domain at the N-terminal not found in other lipases. To understand the influence of N-terminal GST-C domain on the biochemical and structural features of the native lipase, the deletion of the GST-C domain was carried out. The truncated protein was successfully expressed in E. coli BL21(DE3). The molecular weight of truncated AMS3 lipase was approximately ~45 kDa. The number of truncated AMS3 lipase purification folds was higher than native lipase. Various mono and divalent metal ions increased the activity of the AMS3 lipase. The truncated AMS3 lipase demonstrated a similarly broad temperature range, with the pH profile exhibiting higher activity under alkaline conditions. The purified lipase showed a substrate preference for a long carbon chain substrate. In addition, the enzyme activity in organic solvents was enhanced, especially for toluene, Dimethylsulfoxide (DMSO), chloroform and xylene. Molecular simulation revealed that the truncated lipase had increased structural compactness and rigidity as compared to native lipase. Removal of the N terminal GST-C generally improved the lipase biochemical characteristics. This enzyme may be utilized for industrial purposes. Text Antarc* Antarctic MDPI Open Access Publishing Antarctic International Journal of Molecular Sciences 19 2 560 |
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Open Polar |
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MDPI Open Access Publishing |
op_collection_id |
ftmdpi |
language |
English |
topic |
microbial enzyme N-terminal domain biochemical characterization in silico lipase antarctic |
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microbial enzyme N-terminal domain biochemical characterization in silico lipase antarctic Wahhida Latip Raja Raja Abd Rahman Adam Leow Fairolniza Mohd Shariff Nor Kamarudin Mohd Mohamad Ali The Effect of N-Terminal Domain Removal towards the Biochemical and Structural Features of a Thermotolerant Lipase from an Antarctic Pseudomonas sp. Strain AMS3 |
topic_facet |
microbial enzyme N-terminal domain biochemical characterization in silico lipase antarctic |
description |
Lipase plays an important role in industrial and biotechnological applications. Lipases have been subject to modification at the N and C terminals, allowing better understanding of lipase stability and the discovery of novel properties. A thermotolerant lipase has been isolated from Antarctic Pseudomonas sp. The purified Antarctic AMS3 lipase (native) was found to be stable across a broad range of temperatures and pH levels. The lipase has a partial Glutathione-S-transferase type C (GST-C) domain at the N-terminal not found in other lipases. To understand the influence of N-terminal GST-C domain on the biochemical and structural features of the native lipase, the deletion of the GST-C domain was carried out. The truncated protein was successfully expressed in E. coli BL21(DE3). The molecular weight of truncated AMS3 lipase was approximately ~45 kDa. The number of truncated AMS3 lipase purification folds was higher than native lipase. Various mono and divalent metal ions increased the activity of the AMS3 lipase. The truncated AMS3 lipase demonstrated a similarly broad temperature range, with the pH profile exhibiting higher activity under alkaline conditions. The purified lipase showed a substrate preference for a long carbon chain substrate. In addition, the enzyme activity in organic solvents was enhanced, especially for toluene, Dimethylsulfoxide (DMSO), chloroform and xylene. Molecular simulation revealed that the truncated lipase had increased structural compactness and rigidity as compared to native lipase. Removal of the N terminal GST-C generally improved the lipase biochemical characteristics. This enzyme may be utilized for industrial purposes. |
format |
Text |
author |
Wahhida Latip Raja Raja Abd Rahman Adam Leow Fairolniza Mohd Shariff Nor Kamarudin Mohd Mohamad Ali |
author_facet |
Wahhida Latip Raja Raja Abd Rahman Adam Leow Fairolniza Mohd Shariff Nor Kamarudin Mohd Mohamad Ali |
author_sort |
Wahhida Latip |
title |
The Effect of N-Terminal Domain Removal towards the Biochemical and Structural Features of a Thermotolerant Lipase from an Antarctic Pseudomonas sp. Strain AMS3 |
title_short |
The Effect of N-Terminal Domain Removal towards the Biochemical and Structural Features of a Thermotolerant Lipase from an Antarctic Pseudomonas sp. Strain AMS3 |
title_full |
The Effect of N-Terminal Domain Removal towards the Biochemical and Structural Features of a Thermotolerant Lipase from an Antarctic Pseudomonas sp. Strain AMS3 |
title_fullStr |
The Effect of N-Terminal Domain Removal towards the Biochemical and Structural Features of a Thermotolerant Lipase from an Antarctic Pseudomonas sp. Strain AMS3 |
title_full_unstemmed |
The Effect of N-Terminal Domain Removal towards the Biochemical and Structural Features of a Thermotolerant Lipase from an Antarctic Pseudomonas sp. Strain AMS3 |
title_sort |
effect of n-terminal domain removal towards the biochemical and structural features of a thermotolerant lipase from an antarctic pseudomonas sp. strain ams3 |
publisher |
Multidisciplinary Digital Publishing Institute |
publishDate |
2018 |
url |
https://doi.org/10.3390/ijms19020560 |
op_coverage |
agris |
geographic |
Antarctic |
geographic_facet |
Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
International Journal of Molecular Sciences; Volume 19; Issue 2; Pages: 560 |
op_relation |
Biochemistry https://dx.doi.org/10.3390/ijms19020560 |
op_rights |
https://creativecommons.org/licenses/by/4.0/ |
op_doi |
https://doi.org/10.3390/ijms19020560 |
container_title |
International Journal of Molecular Sciences |
container_volume |
19 |
container_issue |
2 |
container_start_page |
560 |
_version_ |
1774724905822060544 |