Catalytic Performance in Nitroarene Reduction of Nanocatalyst Based on Noble Metal Nanoparticles Supported on Polymer/s-Layer Protein Hybrids.

We present a novel bionanocatalyst fabricated by the adsorption-reduction of metal ions on a polyurethane/S-layer protein biotemplate. The bioinspired support was obtained by the adsorption of S-layer proteins (isolated from Lentilactobacillus kefiri) on polyurethane particles. Silver and platinum nanoparticles were well-loaded on the surface of the support after the combination with metallic salts and reduction with H2 at room temperature. Transmission electron microscopy analysis revealed the strawberry-like morphology of the bionanocatalysts with a particle size, dn, of 2.39 nm for platinum and 9.60 nm for silver. Both systems catalyzed the hydrogenation of p-nitrophenol to p-aminophenol with high efficiency in water at mild conditions in the presence of NaBH4. Three different amounts of bionanocatalyst were tested, and in all cases, conversions between 97 and 99% were observed. The catalysts displayed excellent recyclability over ten cycles, and no extensive damage in their nanostructure was noted after them. The bionanocatalysts were stable during their production, storage, and use, thanks to the fact that the biosupport provides an effective driving force in the formation and stabilization of the metallic nanoparticles. The successful bioinspired production strategy and the good catalytic ability of the systems are encouraging in the search for nontoxic, simple, clean, and eco-friendly procedures for the synthesis and exploitation of nanostructures.

Synthesis and Catalytic Application of Silver Nanoparticles Supported on Lactobacillus kefiri S-Layer Proteins.

Research on nanoparticles obtained on biological supports is a topic of growing interest in nanoscience, especially regarding catalytic applications. Silver nanoparticles (AgNPs) have been studied due to their low toxicity, but they tend to aggregation, oxidation, and low stability. In this work, we synthesized and characterized AgNPs supported on S-layer proteins (SLPs) as bidimensional regularly arranged biotemplates. By different reduction strategies, six AgNPs of variable sizes were obtained on two different SLPs. Transmission electron microscopy (TEM) images showed that SLPs are mostly decorated by evenly distributed AgNPs; however, a drastic reduction by NaBH4 led to large AgNPs whereas a smooth reduction with H2 or H2/NaBH4 at low concentration leads to smaller AgNPs, regardless of the SLP used as support. All the nanosystems showed conversion values between 75-80% of p-nitrophenol to p-aminophenol, however, the increment in the AgNPs size led to a great decrease in Kapp showing the influence of reduction strategy in the performance of the catalysts. Density functional theory (DFT) calculations indicated that the adsorption of p-nitrophenolate species through the nitro group is the most favored mechanism, leading to p-aminophenol as the only feasible product of the reaction, which was corroborated experimentally.

Platinum Nanoparticles Obtained at Mild Conditions on S-Layer Protein/Polymer Particle Supports.

This work presents the synthesis of platinum nanoparticles supported on S-layer protein/polymeric particle systems, obtained by combining proteins isolated from Lactobacillus kefiri and an aqueous dispersion of acrylic particles. FTIR spectra of the protein/polymer supports did not show changes in the Amide I band of the proteins, suggesting that proteins maintained their conformation after adsorption. The SAXS spectra and DLS results are consistent with the formation of a protein corona around the polymer particles. After combining the supports with the platinum complex and subsequently reducing the combination with hydrogen at mild conditions, we obtained colloidal nanocomposite materials. In these, platinum nanoparticles with diameters around 3 nm located on the surface of the protein/polymer supports were observed by TEM. The obtained nanosystems showed catalytic activity in the reduction of p-nitrophenol with NaBH4 at room temperature with conversions of 100% for reaction times of 50 to 70 min.

Protective effect of a mixture of kefir-isolated lactic acid bacteria and yeasts in a hamster model of Clostridium difficile infection.

The objective of this work was to test the protective effect of a mixture (MM) constituted by kefir-isolated microorganisms (Lactobacillus plantarum, Lactobacillus kefir, Lc. lactis, Kluyveromyces marxianus and Saccharomyces cerevisiae) in a hamster model of infection with Clostridium difficile, an anaerobic Gram-positive bacterium that causes diarrhoea. Placebo or MM was administered ad libitum in drinking water from day 0 to the end of treatment. Hamsters received orally 200 µg of clyndamicin at day 7 and then were infected with 1 × 10(8) CFU of C. difficile by gavage. Development of diarrhoea and death was registered until the end of the protocol. Surviving animals were sacrificed at day 16, and a test for biological activity of clostridial toxins and histological stainings were performed in caecum samples. Six of seven infected animals developed diarrhoea and 5/7 died at the end of the experimental protocol. The histological sections showed oedema and inflammatory infiltrates with neutrophils and crypt abscesses. In the group of animals infected and treated with MM1/1000, only 1 of 7 hamsters showed diarrhoea and none of them died. The histological sections showed only a slight thickening of the mucosa with presence of lymphocytic infiltrate. These results demonstrate that an oral treatment with a mixture of kefir-isolated bacteria and yeasts was able to prevent diarrhoea and enterocolitis triggered by C. difficile.

Effect of freeze-drying on viability and in vitro probiotic properties of a mixture of lactic acid bacteria and yeasts isolated from kefir.

The effect of freeze-drying on viability and probiotic properties of a microbial mixture containing selected bacterial and yeast strains isolated from kefir grains (Lactobacillus kefir, Lactobacillus plantarum, Lactococcus lactis, Saccharomyces cerevisiae and Kluyveromyces marxianus) was studied. The microorganisms were selected according to their potentially probiotic properties in vitro already reported. Two types of formulations were performed, a microbial mixture (MM) suspended in milk and a milk product fermented with MM (FMM). To test the effect of storage on viability of microorganisms, MM and FMM were freeze-dried and maintained at 4°C for six months. After 180 days of storage at 4°C, freeze-dried MM showed better survival rates for each strain than freeze-dried FMM. The addition of sugars (trehalose or sucrose) did not improve the survival rates of any of the microorganisms after freeze-drying. Freeze-drying did not affect the capacity of MM to inhibit growth of Shigella sonnei in vitro, since the co-incubation of this pathogen with freeze-dried MM produced a decrease of 2 log in Shigella viability. The safety of freeze-dried MM was tested in mice and non-translocation of microorganisms to liver or spleen was observed in BALB/c mice feed ad libitum during 7 or 20 days. To our knowledge, this is the first report about the effect of freeze-drying on viability, in vitro probiotic properties and microbial translocation of a mixture containing different strains of both bacteria and yeasts isolated from kefir.