Prospecting Potential Inhibitors of Sortase A from Enterococcus faecalis: A Multidrug Resistant Bacteria, through In-silico and In-vitro Approaches.

BACKGROUND: Enterococcus faecalis (Ef) infections are becoming dreadfully common in hospital environments. Infections caused by Ef are difficult to treat because of its acquired resistance to different class of antibiotics, making it a multidrug resistant bacteria. Key pathogenic factor of Ef includes its ability to form biofilm on the surface of diagnostic and other medical devices. Sortase A (SrtA) is a cysteine transpeptidase which plays a pivotal role in the formation of biofilm in Ef, hence, it is considered as an important enzyme for the pathogenesis of Ef. Thus, inhibition of (SrtA) will affect biofilm formation, which will reduce its virulence and eventually Ef infection will be abridged. OBJECTIVE: To find potential inhibitors of Enterococcus faecalis Sortase A (EfSrtA) through insilico and in-vitro methods. METHODS: Gene coding for EfSrtA was cloned, expressed and purified. Three-dimensional model of EfSrtA was created using Swiss-Model workspace. In-silico docking studies using Autodock vina and molecular dynamics simulations of the modelled structures using Gromacs platform were performed to explore potential lead compounds against EfSrtA. In-vitro binding experiments using spectrofluorometric technique was carried out to confirm and validate the study. RESULTS: In-silico docking and in-vitro binding experiments revealed that curcumin, berberine and myricetin bound to EfSrtA at nanomolar concentrations with high affinity. CONCLUSION: This is a first structural report of EfSrtA with curcumin, berberine and myricetin. Taking in account the herbal nature of these compounds, the use of these compounds as inhibitors will be advantageous. This study validated curcumin, berberine and myricetin as potential inhibitors of EfSrtA.

Transcriptional profiling and genes involved in acquired thermotolerance in Banana: a non-model crop.

Banana is a non- model crop plant, and one of the most important crops in the tropics and sub tropics. Heat stress is the major abiotic stress affecting banana crop production because of its long growth period and is likely to become a threat due to global warming. To understand an acquired thermotolerance phenomenon at the molecular level, the RNA-seq approach was employed by adapting TIR method. A total of 136.38 million high quality reads were assembled. Differentially expressed genes under induction (I) was 3936, I + L was 2268 and lethal stress was 907 compared to control. Gene ontology and DGE analysis showed that genes related to heat shock factors, heat shock proteins, stress associated proteins, ROS scavenging, fatty acid metabolism, protein modification were significantly up regulated during induction, thus preparing the organism or tissue at molecular and cellular level for acquired thermotolerance. KEGG pathway analysis revealed the significant enrichment of pathways involved in protein processing, MAPK signaling and HSPs which indicates that these processes are conserved and involved in thermo tolerance. Thus, this study provides insights into the acquired thermotolerance phenomena in plants especially banana.