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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ftdoajarticles:oai:doaj.org/article:8a91a98fd3394490953ce3051adc1153 2023-05-15T13:43:17+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 Noor Zaliha Raja Abd Rahman Adam Thean Chor Leow Fairolniza Mohd Shariff Nor Hafizah Ahmad Kamarudin Mohd Shukuri Mohamad Ali 2018-02-01T00:00:00Z https://doi.org/10.3390/ijms19020560 https://doaj.org/article/8a91a98fd3394490953ce3051adc1153 EN eng MDPI AG http://www.mdpi.com/1422-0067/19/2/560 https://doaj.org/toc/1422-0067 1422-0067 doi:10.3390/ijms19020560 https://doaj.org/article/8a91a98fd3394490953ce3051adc1153 International Journal of Molecular Sciences, Vol 19, Iss 2, p 560 (2018) microbial enzyme N-terminal domain biochemical characterization in silico lipase antarctic Biology (General) QH301-705.5 Chemistry QD1-999 article 2018 ftdoajarticles https://doi.org/10.3390/ijms19020560 2022-12-31T00:30:58Z 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. Article in Journal/Newspaper Antarc* Antarctic Directory of Open Access Journals: DOAJ Articles Antarctic International Journal of Molecular Sciences 19 2 560 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
microbial enzyme N-terminal domain biochemical characterization in silico lipase antarctic Biology (General) QH301-705.5 Chemistry QD1-999 |
spellingShingle |
microbial enzyme N-terminal domain biochemical characterization in silico lipase antarctic Biology (General) QH301-705.5 Chemistry QD1-999 Wahhida Latip Raja Noor Zaliha Raja Abd Rahman Adam Thean Chor Leow Fairolniza Mohd Shariff Nor Hafizah Ahmad Kamarudin Mohd Shukuri 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 Biology (General) QH301-705.5 Chemistry QD1-999 |
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 |
Article in Journal/Newspaper |
author |
Wahhida Latip Raja Noor Zaliha Raja Abd Rahman Adam Thean Chor Leow Fairolniza Mohd Shariff Nor Hafizah Ahmad Kamarudin Mohd Shukuri Mohamad Ali |
author_facet |
Wahhida Latip Raja Noor Zaliha Raja Abd Rahman Adam Thean Chor Leow Fairolniza Mohd Shariff Nor Hafizah Ahmad Kamarudin Mohd Shukuri 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 |
MDPI AG |
publishDate |
2018 |
url |
https://doi.org/10.3390/ijms19020560 https://doaj.org/article/8a91a98fd3394490953ce3051adc1153 |
geographic |
Antarctic |
geographic_facet |
Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
International Journal of Molecular Sciences, Vol 19, Iss 2, p 560 (2018) |
op_relation |
http://www.mdpi.com/1422-0067/19/2/560 https://doaj.org/toc/1422-0067 1422-0067 doi:10.3390/ijms19020560 https://doaj.org/article/8a91a98fd3394490953ce3051adc1153 |
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_ |
1766186672085532672 |