Genomic and metabolic properties of Staphylococcus gallinarum FCW1 MCC4687 isolated from naturally fermented coconut water towards GRAS assessment.

Staphylococcus gallinarum FCW1 was isolated from naturally fermented coconut-water and identified by biochemical and molecular methods. Probiotic characterization and safety assessment were conducted through a series of in vitro tests. A high survival rate was observed when the strain was tested for resistance to bile, lysozyme, simulated gastric and intestinal fluid, phenol, and different temperature and salt concentrations. The strain showed antagonism against some pathogens, was susceptible to all antibiotics tested except penicillin, and showed no hemolytic and DNase activity. Hydrophobicity, autoaggregation, biofilm formation, and antioxidation tests indicated that the strain possessed a high adhesive and antioxidant ability. Enzymatic activity was used to evaluate the metabolic capacities of the strain. In-vivo experiment on zebrafish was performed to check its safety status. The whole-genome sequencing indicated that the genome contained 2,880,305 bp with a GC content of 33.23%. The genome annotation confirmed the presence of probiotic-associated genes and genes for oxalate degradation, sulfate reduction, acetate metabolism, and ammonium transport in the FCW1 strain, adding to the theory that this strain may be helpful in treating kidney stones. This study revealed that the strain FCW1 might be an excellent potential probiotic in developing fermented coconut beverages and treating and preventing kidney stone disease.

Purification of WAP domain-containing antimicrobial peptides from green tiger shrimp Peaneaus semisulcatus.

Antimicrobial peptide crustin was isolated and purified from Penaeus semisulcatus using Sephadox G-100 column gel filtration chromatography. P. semisulcatus crustins was observed as a single band with 14 kDa of molecular weight on SDS-PAGE and the retention time of 46 min in RP-HPLC. Circular dichroism spectra of P. semisulcatus crustin showed alpha helices in its secondary structure followed by random coils. Crystalline nature and functional groups arrangement were investigated by X-Ray Diffraction (XRD) and Fourier Transform Infra-Red spectroscopy (FTIR). P. semisulcatus crustin showed the effective antibacterial activity against Gram positive strains B. thuringienisis (4 µg/ml) and B. pumilis (6 µg/ml) when compare to Gram negative strains. Biofilm Inhibitory Concentration (BIC) were determined for these strains and percentage of biofilm inhibition was confirmed and visualized through in sit microscopic analysis. Hence, we reported the effect of crustin on biofilm inhibition and eradication at low concentrations by using crystal violet staining and confocal microscopic observations. In addition, haemolytic activity of this purified crustin also analysed using human RBCs. The results of this study, suggests that this bio peptide crustin is a potential and promising therapeutic agent to treat drug resistant bacteria and biofilm-related infections.