Data_Sheet_1_Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase Family.docx

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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Bibliographic Details
Main Authors: Xiaoyu Liu, Mingyang Zhou, Rui Sun, Shu Xing, Tao Wu, Hailun He, Jianbin Chen, John Kevin Bielicki
Format: Dataset
Language:unknown
Published: 2022
Subjects:
Microbiology
Microbial Genetics
Microbial Ecology
Mycology
esterase
Antarctic
bacterium
soil
new esterase family XXI
Antarc*
Online Access:https://doi.org/10.3389/fmicb.2022.855658.s001
https://figshare.com/articles/dataset/Data_Sheet_1_Characterization_of_a_Novel_Esterase_Est33_From_an_Antarctic_Bacterium_A_Representative_of_a_New_Esterase_Family_docx/19777459
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Summary: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.

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