Back to Search
Start Over
Biochemical characterization of the endo-α-N-acetylgalactosaminidase pool of the human gut symbiont Tyzzerella nexilis.
- Source :
-
Carbohydrate research [Carbohydr Res] 2020 Apr; Vol. 490, pp. 107962. Date of Electronic Publication: 2020 Feb 26. - Publication Year :
- 2020
-
Abstract
- Three large (2084-, 984-, and 2104-amino acids) endo-α-N-acetylgalactosaminidase candidate genes from the commensal human gut bacterium Tyzzerella nexilis were successfully cloned and subsequently expressed in Escherichia coli. Activity tests of the purified proteins revealed that two of the candidate genes (Tn0153 and Tn2105) were able to hydrolyze the disaccharide unit from Galβ1-3GalNAc-α-pNP. The biochemical characterization revealed optimum pH conditions of 4.0 for both enzymes and temperature optima of 50 °C. The addition of 2-mercaptoethanol, Triton X-100 and urea had only minor effects on the activity of the enzymes, and the addition of imidazole and sodium dodecyl sulfate led to a significant reduction of the enzymes' activities. A mutational study identified and confirmed the role of the catalytically significant amino acids. The present study describes the first functional characterization of members of the GH101 family from this human gut symbiont.<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 © 2020 Elsevier Ltd. All rights reserved.)
- Subjects :
- Bacterial Proteins
Clostridiales enzymology
Disaccharides metabolism
Escherichia coli genetics
Escherichia coli growth & development
Humans
Hydrolysis
Mercaptoethanol pharmacology
Mutation
Octoxynol pharmacology
Substrate Specificity
Symbiosis
Urea pharmacology
Cloning, Molecular methods
Clostridiales physiology
Gastrointestinal Tract microbiology
alpha-N-Acetylgalactosaminidase genetics
alpha-N-Acetylgalactosaminidase metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1873-426X
- Volume :
- 490
- Database :
- MEDLINE
- Journal :
- Carbohydrate research
- Publication Type :
- Academic Journal
- Accession number :
- 32169671
- Full Text :
- https://doi.org/10.1016/j.carres.2020.107962