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Selection and characterization of alanine racemase inhibitors against Aeromonas hydrophila.
- Source :
-
BMC microbiology [BMC Microbiol] 2017 May 25; Vol. 17 (1), pp. 122. Date of Electronic Publication: 2017 May 25. - Publication Year :
- 2017
-
Abstract
- Background: Combining experimental and computational screening methods has been of keen interest in drug discovery. In the present study, we developed an efficient screening method that has been used to screen 2100 small-molecule compounds for alanine racemase Alr-2 inhibitors.<br />Results: We identified ten novel non-substrate Alr-2 inhibitors, of which patulin, homogentisic acid, and hydroquinone were active against Aeromonas hydrophila. The compounds were found to be capable of inhibiting Alr-2 to different extents with 50% inhibitory concentrations (IC <subscript>50</subscript> ) ranging from 6.6 to 17.7 μM. These compounds inhibited the growth of A. hydrophila with minimal inhibitory concentrations (MICs) ranging from 20 to 120 μg/ml. These compounds have no activity on horseradish peroxidase and D-amino acid oxidase at a concentration of 50 μM. The MTT assay revealed that homogentisic acid and hydroquinone have minimal cytotoxicity against mammalian cells. The kinetic studies indicated a competitive inhibition of homogentisic acid against Alr-2 with an inhibition constant (K <subscript>i</subscript> ) of 51.7 μM, while hydroquinone was a noncompetitive inhibitor with a K <subscript>i</subscript> of 212 μM. Molecular docking studies suggested that homogentisic acid binds to the active site of racemase, while hydroquinone lies near the active center of alanine racemase.<br />Conclusions: Our findings suggested that combining experimental and computational methods could be used for an efficient, large-scale screening of alanine racemase inhibitors against A. hydrophila that could be applied in the development of new antibiotics against A. hydrophila.
- Subjects :
- Aeromonas hydrophila enzymology
Aeromonas hydrophila growth & development
Anti-Bacterial Agents chemistry
Catalytic Domain drug effects
Cell Survival drug effects
D-Amino-Acid Oxidase drug effects
Drug Evaluation, Preclinical
Enzyme Assays
HeLa Cells drug effects
Homogentisic Acid antagonists & inhibitors
Homogentisic Acid chemistry
Horseradish Peroxidase drug effects
Humans
Hydroquinones antagonists & inhibitors
Hydroquinones chemistry
Inhibitory Concentration 50
Kinetics
Microbial Sensitivity Tests
Molecular Docking Simulation methods
Patulin antagonists & inhibitors
Patulin chemistry
Aeromonas hydrophila drug effects
Alanine Racemase drug effects
Anti-Bacterial Agents pharmacology
Drug Discovery
Subjects
Details
- Language :
- English
- ISSN :
- 1471-2180
- Volume :
- 17
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- BMC microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 28545531
- Full Text :
- https://doi.org/10.1186/s12866-017-1010-x