An in vitro and in silico identification of antibiofilm small molecules from seawater metaclone SWMC166 against Vibrio cholerae O1
| Autor: | Shunmugiah Karutha Pandian, Murugan Rajalaxmi, Rajamohamed Beema Shafreen, Karuppiah Chithiraiselvi |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
In silico 030106 microbiology Virulence Biology medicine.disease_cause Small Molecule Libraries 03 medical and health sciences medicine Computer Simulation Seawater Molecular Biology Extracellular Polymeric Substance Matrix Vibrio cholerae O1 Biofilm Quorum Sensing Gene Expression Regulation Bacterial Cell Biology In vitro Molecular Docking Simulation Quorum sensing 030104 developmental biology Subcloning Biochemistry Vibrio cholerae Docking (molecular) Biofilms Metagenome Thermodynamics Hydrophobic and Hydrophilic Interactions Peptide Hydrolases |
| Zdroj: | Molecular and Cellular Probes. 39:14-24 |
| ISSN: | 0890-8508 |
| Popis: | This study aimed to determine the antibiofilm activity of seawater microbes against Vibrio cholerae (VCO1) through functional metagenomics approach. A metagenomic library was constructed from Palk Bay seawater and the library was screened to identify the biofilm inhibitory metaclone. Metaclone SWMC166 (harbouring ∼30 kb metagenomic insert) was found to exhibit antibiofilm activity against VCO1. The biofilm inhibitory potential of partially purified ethyl acetate extract of SWMC166 (EA166) was further evaluated through microscopic studies and biochemical assays. Further, EA166 treated VCO1 divulged up-regulation of genes involved in high cell density-mediated quorum sensing (QS) pathway which was analysed by real-time PCR. In order to identify the genes of interest (within ∼30 kb insert), subcloning was performed through shotgun approach. Small molecules from positive subclones SC5 and SC8 were identified through HRLC-MS analysis. Resulted small molecules were docked against QS receptors of V. cholerae to identify the bioactive metabolites. Docking studies revealed that totally seven metabolites were able to interact with QS receptors that can possibly trigger the QS cascade and sequentially inhibit the biofilm formation and virulence factors of VCO1. |
| Databáze: | OpenAIRE |
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