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Exploration of novel trehalases from cold-adapted Variovorax sp. PAMC28711: Functional characterization.
- Source :
-
International journal of biological macromolecules [Int J Biol Macromol] 2024 Jun; Vol. 271 (Pt 1), pp. 132503. Date of Electronic Publication: 2024 May 18. - Publication Year :
- 2024
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Abstract
- The cold-adapted bacterium Variovorax sp. PAMC28711 possesses two distinct glycoside hydrolase (GH) families of trehalase, GH15 and GH37. While numerous studies have explored bacterial trehalase, the presence of two different trehalase genes within a single strain has not been reported until now. Interestingly, despite both GH37 and GH15 trehalases serving the same purpose of degrading trehalose, but do not share the sequence similarity. The substrate specificity assay confirmed that Vtre37 and Vtre15 displayed hydrolytic activity on α, α-trehalose. The key catalytic sites were identified as D280 and E469 in Vtre37 and E389 and E554 in Vtre15 through site-directed mutation and confirmed these two enzymes belong to trehalase. In addition, Vtre37 exhibited a relatively high level of enzyme activity of 1306.33 (±53.091) μmolmg <superscript>-1</superscript> , whereas Vtre15 showed enzyme activity of 408.39 (±12.503) μmolmg <superscript>-1</superscript> . Moreover, Vtre37 performed admirably showing resistance to ethanol (10 %), with high stable at acidic pH range. Furthermore, both prediction and experimental results indicate that validoxylamine A showed a potent inhibitory activity against Vtre37 trehalase with a K <subscript>i</subscript> value of 16.85 nM. Therefore, we postulate that Vtre37 could be utilized as an ethanol enhancer and designed for screening inhibitors related to the trehalose degradation pathway. Additionally, we believe that characterizing these bacterial trehalase contributes to a better understanding of trehalose metabolism and its biological importance in bacteria.<br />Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.<br /> (Copyright © 2023. Published by Elsevier B.V.)
- Subjects :
- Substrate Specificity
Catalytic Domain
Trehalose metabolism
Trehalose pharmacology
Hydrogen-Ion Concentration
Hydrolysis
Kinetics
Bacterial Proteins metabolism
Bacterial Proteins genetics
Bacterial Proteins chemistry
Amino Acid Sequence
Enzyme Stability
Adaptation, Physiological
Trehalase metabolism
Trehalase genetics
Trehalase chemistry
Cold Temperature
Comamonadaceae enzymology
Comamonadaceae genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1879-0003
- Volume :
- 271
- Issue :
- Pt 1
- Database :
- MEDLINE
- Journal :
- International journal of biological macromolecules
- Publication Type :
- Academic Journal
- Accession number :
- 38768913
- Full Text :
- https://doi.org/10.1016/j.ijbiomac.2024.132503