Fabrication and characterisation of human gut microbiome derived exopolysaccharide mediated silver nanoparticles - An in-vitro and in-vivo approach of Bio-Pm-AgNPs targeting Vibrio cholerae

Utilising bacteria to produce silver nanoparticles was highly favoured due to its ability to minimise costs and mitigate any potential negative environmental impact. Exopolysaccharides (EPS) extracted from the human gut microbe have demonstrated remarkable efficacy in combating various bacterial infections. Exopolysaccharide (EPS), a naturally occurring biomolecule found in the human gut isolate Proteus mirabilis DMTMMR-11, was characterised using analytical techniques such as Fourier transform infrared spectroscopy (FTIR), H-1-nuclear magnetic resonance, C-13-nuclear magnetic resonance (NMR), and chemical composition analysis, which confirms the presence of carbohydrates (81.03 +/- 0.23), proteins (4.22 +/- 1.2), uronic acid (12.1 +/- 0.12), and nucleic acid content (2.44 +/- 0.15) in exopolysaccharide. The one factor at a time (OFAT) and response surface methodology (RSM) - central composite design (CCD) approaches were used to optimise the production of Bio-Pm-AgNPs, leading to an increase in yield of up to 1.86 g/l. The Bio-Pm-AgNPs were then subjected to Fourier transform infrared spectroscopy (FTIR) which determines the functional groups, X-ray diffractometer confers that Bio-Pm-AgNPs are crystalline in nature, field emission-scanning electron microscopy (FE-SEM) reveals the morphology of Bio-Pm-AgNPs, energy dispersive X-ray spectroscopy (EDX) confirms the presence of elements like Ag, C and O, high-resolution transmission electron microscopy (HR-TEM) determines that the Bio-Pm-AgNPs are sphere-shaped at 75 nm. Dynamic light scattering (DLS) and zeta potential analysis were also carried out to reveal the physiological nature of Bio-Pm-AgNPs. Bio-Pm-AgNPs have a promising effect on the inhibitory mechanism of Vibrio cholerae cells at a MIC concentration of 20 mu g/ml which significantly affects cellular respiration and energy metabolism through glycolysis and TCA cycles by deteriorating the enzyme responsible for ATP and NADH utilisation. The action of Bio-Pm-AgNPs reduces the purity and concentration of nucleic acids, which leads to higher DNA damage. In-vivo analysis reveals that the treatment of Bio-Pm-AgNPs decreased the colonisation of V. cholerae and improved the survival rates in C. elegans.