Exopolysaccharide from Lactococcus hircilactis CH4 and Lactobacillus delbrueckii GRIPUMSK as new therapeutics to treat biofilm pathogens, oxidative stress and human colon adenocarcinoma.

Naturally occurring biopolymers like exopolysaccharides (EPS) secreted by lactic acid bacteria (LAB) has gained significant attention as they are cost effective, renewable and safe. In order to prevent the rapid increase in antibiotic resistant bacteria, the EPS of LAB offers novel approach of targeting the antibiotic resistant pathogens by limiting their effects on environment. Accordingly, in this study, the production, purification, characterization and biological properties of exopolysaccharides from Lactococcus hircilactis strain CH4 and Lactobacillus delbrueckii strain GRIPUMSK were performed. The optimization of lactic acid bacterial strains for exopolysaccharide production was done by response surface methodology and changing the carbon sources in the growth media. The carbohydrate and protein of exopolysaccharide 1 were 79.7 % and 8.7 % respectively and exopolysaccharide 2 were 75.2 % and 9.3 % respectively. When compared with the commercial emulsifier sodium dodecyl sulfate, both the exopolysaccharides have shown good emulsifying activity. Both the exopolysaccharides were linear homo-polysaccharide as determined by Fourier transform infrared spectroscopy and Nuclear magnetic resonance spectra. Scanning electron microscopy showed that the exopolysaccharides were porous and capable of holding water. The exopolysaccharides were partially crystalline as confirmed by X-ray diffraction spectra. Exopolysaccharides from L. hircilactis and L. delbrueckii exhibited significant antimicrobial activity against H. pylori, S. flexneri, S. pyogenes, E. faecalis and C. albicans. Both the exopolysaccharides revealed significant 2,2-diphenyl-1-picrylhydrazyl and hydrogen peroxide scavenging ability with the IC50 value of 100 µg/ml and 80 µg/ml respectively. Exopolysaccharides from L. hircilactis and L. delbrueckii at 100 µg/ml showed significant anticancer activity on HT-29 cells with 58.4 % and 58.7 % respectively. These findings proved that exopolysaccharides from the two selected lactic acid bacterial strains could be explored as natural bioactive carbohydrate polymer for biomedical applications.

Nanomedicine for drug resistant pathogens and COVID-19 using mushroom nanocomposite inspired with bacteriocin - A review.

Multidrug resistant (MDR) pathogens have become a major global health challenge and have severely threatened the health of society. Current conditions have become worse as a result of the COVID-19 pandemic, and infection rates in the future will rise. It is necessary to design, respond effectively, and take action to address these challenges by investigating new avenues. In this regard, the fabrication of metal NPs utilized by various methods, including green synthesis using mushroom, is highly versatile, cost-effective, eco-compatible, and superior. In contrast, biofabrication of metal NPs can be employed as a powerful weapon against MDR pathogens and have immense biomedical applications. In addition, the advancement in nanotechnology has made possible to modify the nanomaterials and enhance their activities. Metal NPs with biomolecules composite prevent the microbial adhesion and kills the microbial pathogens through biofilm formation. Bacteriocin is an excellent antimicrobial peptide that works well as an augmentation substance to boost the antimicrobial effects. As a result, we concentrate on the creation of new, eco-compatible mycosynthesized metal NPs with bacteriocin nanocomposite via electrostatic, covalent, or non-covalent bindings. The synergistic benefits of metal NPs with bacteriocin to combat MDR pathogens and COVID-19, as well as other biomedical applications, are discussed in this review. Moreover, the importance of the adverse outcome pathway (AOP) in risk analysis of manufactured metal nanocomposite nanomaterial and their future possibilities were also discussed.

Antimicrobial, anti-biofilm, antioxidant and cytotoxic effects of bacteriocin by Lactococcus lactis strain CH3 isolated from fermented dairy products-An in vitro and in silico approach.

The current study aimed to screen bacteriocin producing LAB from different dairy products and evaluation of their biological properties. Initially, 12 (4-chess, 4-curd, and 4-yohurt) LAB species were isolated and only 4 isolates alone were selected based on their clear yellow halo zone around the colonies in the selective medium. The selected 4 isolates were identified based on their morphological and biochemical characteristics. Among them, the strain CH3 have showed better antimicrobial effects on selected human pathogens. The isolated strain CH3 were further identified as Lactococcus lactis strain CH3 (MZ636710) by SEM imaging and 16 s rRNA molecular sequencing. Bacteriocin was extracted from L. lactis strain CH3 and partially purified using 60 % ammonium sulphate and then completely purified by G-50 column chromatography. The purified bacteriocin showed a specific activity of 5859.37 AU/mg in 24.7 % of recovery and 10.9-fold purification. The molecular weight of bacteriocin was 3.5 kDa as observed in SDS-PAGE. The bacteriocin showed sensitivity to proteolytic enzymes and resistance to high temperature, wide range of pH, organic solvents and detergents. FT-IR spectral studies of bacteriocin detected the existence of OH/NH-stretching, CH, and COC and CO bonds. NMR spectrum showed one doublet and 4 various singlet peaks at different ppm, indicating the occurrence of six amino acids in the structure of purified bacteriocin. The purified bacteriocin have shown stronger antimicrobial and anti-biofilm activity against selected human pathogens at 100 µg/mL. SEM showed the evidence of structural deformation and loss of membrane integrity of bacterial cells treated with bacteriocin. Bacteriocin exhibited greater DPPH radical scavenging potential with an EC50 value of 12.5 µg/mL. Bacteriocin have not shown significant toxicity on normal human dermal fibroblast (NHDF) cells (83.2 % at 100 µg/ mL). Furthermore, in silico studies using molecular modeling and docking were performed to know the proteins involved in antimicrobial action. The results suggests that bacteriocin could be an alternative to combat AMR pathogens and more suitable for food and dairy industries to preserve food without contamination.