Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase Family
Studies of microorganisms from extreme environments can sometimes reveal novel proteins with unique properties. Here, we identified a novel esterase gene (Est33) from an Antarctic bacterium. The protein was expressed and purified for biochemical characterizations. Site-mutation variants including S9...
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ftpubmed:oai:pubmedcentral.nih.gov:9152352 2023-05-15T13:36:33+02:00 Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase Family Liu, Xiaoyu Zhou, Mingyang Sun, Rui Xing, Shu Wu, Tao He, Hailun Chen, Jianbin Bielicki, John Kevin 2022-05-17 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9152352/ https://doi.org/10.3389/fmicb.2022.855658 en eng Frontiers Media S.A. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9152352/ http://dx.doi.org/10.3389/fmicb.2022.855658 Copyright © 2022 Liu, Zhou, Sun, Xing, Wu, He, Chen and Bielicki. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. CC-BY Front Microbiol Microbiology Text 2022 ftpubmed https://doi.org/10.3389/fmicb.2022.855658 2022-06-05T01:11:39Z Studies of microorganisms from extreme environments can sometimes reveal novel proteins with unique properties. Here, we identified a novel esterase gene (Est33) from an Antarctic bacterium. The protein was expressed and purified for biochemical characterizations. Site-mutation variants including S94A, D205A, and H233A were constructed to explore the structure–function relationship of the catalytic triad of Est33, and we found mutating Ser(94), Asp(205), and His(233) residues lead to a complete loss of enzyme activity. In addition, the catalytic Ser(94) located in a conserved pentapeptide motif GVSWG. Phylogenetic analysis showed that Est33 and its closely related homologs belonged to an independent group apart from other known family members, indicating that Est33 represented a new family of esterase. The Est33 enzyme was found to be a cold-active esterase retaining 25%–100% activity from 10°C to 30°C and to have optimal catalytic activity toward p-nitrophenol acetate (30°C and pH7.5). The serine modifying reagent phenylmethylsulfonyl fluoride inhibited the activity of Est33 by 77.34%, while thiol reagents such as dithiol threitol (DTT) activated the enzyme by 3-fold. Metal chelating reagents EDTA had no effects, indicating that Est33 is not a metalloenzyme. Collectively, these results indicate that Est33 constitutes the first member of a novel esterase family XXI that has been identified. Text Antarc* Antarctic PubMed Central (PMC) Antarctic Frontiers in Microbiology 13 |
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English |
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Microbiology |
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Microbiology Liu, Xiaoyu Zhou, Mingyang Sun, Rui Xing, Shu Wu, Tao He, Hailun Chen, Jianbin Bielicki, John Kevin Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase Family |
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Microbiology |
description |
Studies of microorganisms from extreme environments can sometimes reveal novel proteins with unique properties. Here, we identified a novel esterase gene (Est33) from an Antarctic bacterium. The protein was expressed and purified for biochemical characterizations. Site-mutation variants including S94A, D205A, and H233A were constructed to explore the structure–function relationship of the catalytic triad of Est33, and we found mutating Ser(94), Asp(205), and His(233) residues lead to a complete loss of enzyme activity. In addition, the catalytic Ser(94) located in a conserved pentapeptide motif GVSWG. Phylogenetic analysis showed that Est33 and its closely related homologs belonged to an independent group apart from other known family members, indicating that Est33 represented a new family of esterase. The Est33 enzyme was found to be a cold-active esterase retaining 25%–100% activity from 10°C to 30°C and to have optimal catalytic activity toward p-nitrophenol acetate (30°C and pH7.5). The serine modifying reagent phenylmethylsulfonyl fluoride inhibited the activity of Est33 by 77.34%, while thiol reagents such as dithiol threitol (DTT) activated the enzyme by 3-fold. Metal chelating reagents EDTA had no effects, indicating that Est33 is not a metalloenzyme. Collectively, these results indicate that Est33 constitutes the first member of a novel esterase family XXI that has been identified. |
format |
Text |
author |
Liu, Xiaoyu Zhou, Mingyang Sun, Rui Xing, Shu Wu, Tao He, Hailun Chen, Jianbin Bielicki, John Kevin |
author_facet |
Liu, Xiaoyu Zhou, Mingyang Sun, Rui Xing, Shu Wu, Tao He, Hailun Chen, Jianbin Bielicki, John Kevin |
author_sort |
Liu, Xiaoyu |
title |
Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase Family |
title_short |
Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase Family |
title_full |
Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase Family |
title_fullStr |
Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase Family |
title_full_unstemmed |
Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase Family |
title_sort |
characterization of a novel esterase est33 from an antarctic bacterium: a representative of a new esterase family |
publisher |
Frontiers Media S.A. |
publishDate |
2022 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9152352/ https://doi.org/10.3389/fmicb.2022.855658 |
geographic |
Antarctic |
geographic_facet |
Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
Front Microbiol |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9152352/ http://dx.doi.org/10.3389/fmicb.2022.855658 |
op_rights |
Copyright © 2022 Liu, Zhou, Sun, Xing, Wu, He, Chen and Bielicki. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.3389/fmicb.2022.855658 |
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Frontiers in Microbiology |
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13 |
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1766080328151072768 |