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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Frontiers Media S.A.
2022-05-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2022.855658/full |
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| author | Xiaoyu Liu Mingyang Zhou Rui Sun Shu Xing Tao Wu Hailun He Jianbin Chen Jianbin Chen John Kevin Bielicki |
| author_facet | Xiaoyu Liu Mingyang Zhou Rui Sun Shu Xing Tao Wu Hailun He Jianbin Chen Jianbin Chen John Kevin Bielicki |
| author_sort | Xiaoyu Liu |
| collection | DOAJ |
| 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 Ser94, Asp205, and His233 residues lead to a complete loss of enzyme activity. In addition, the catalytic Ser94 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. |
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| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-12T03:42:02Z |
| publishDate | 2022-05-01 |
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| series | Frontiers in Microbiology |
| spelling | doaj.art-bb56613108e741d0b7c70178b6f6ee102022-12-22T00:39:39ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2022-05-011310.3389/fmicb.2022.855658855658Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase FamilyXiaoyu Liu0Mingyang Zhou1Rui Sun2Shu Xing3Tao Wu4Hailun He5Jianbin Chen6Jianbin Chen7John Kevin Bielicki8School of Chemistry and Chemical Engineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan, ChinaSchool of Chemistry and Chemical Engineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan, ChinaSchool of Chemistry and Chemical Engineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan, ChinaSchool of Chemistry and Chemical Engineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan, ChinaSchool of Chemistry and Chemical Engineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan, ChinaState Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha, ChinaSchool of Chemistry and Chemical Engineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan, ChinaShandong Provincial Key Laboratory of Molecular Engineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan, ChinaLawrence Berkeley National Laboratory, University of California at Berkeley, Berkeley, CA, United StatesStudies 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 Ser94, Asp205, and His233 residues lead to a complete loss of enzyme activity. In addition, the catalytic Ser94 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.https://www.frontiersin.org/articles/10.3389/fmicb.2022.855658/fullesteraseAntarcticbacteriumsoilnew esterase family XXI |
| spellingShingle | Xiaoyu Liu Mingyang Zhou Rui Sun Shu Xing Tao Wu Hailun He Jianbin Chen Jianbin Chen John Kevin Bielicki Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase Family Frontiers in Microbiology esterase Antarctic bacterium soil new esterase family XXI |
| title | 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_short | 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 |
| topic | esterase Antarctic bacterium soil new esterase family XXI |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2022.855658/full |
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