Nanopore-based metagenomic analysis of the impact of nanoparticles on soil microbial communities.

The current trend of using nanotechnology products in all spheres of human life, including for crop improvement may have a possible impact on soil microorganisms which influence soil and plant health. Nanopore-based metagenomic study reported here used full-length 16S rRNA gene sequences to assess shifts in community composition of soil microorganisms when treated with silver, titanium dioxide and zinc oxide nanoparticles (S-NP, T-NP, Z-NP, respectively). Firmicutes and Proteobacteria were the two dominant phyla in this soil, and there were no significant differences (p < 0.05) observed in these phyla across treatments. However, in the phylum Firmicutes, the abundance of the order Clostridiales showed a significant decrease (p < 0.05) in the presence of S-NP. Similarly, in the phylum Proteobacteria, a significant decrease in the presence of S-NP was seen for two orders, Vibrionales (p < 0.05) and Rhodobacterales (p < 0.01). Analysis at a further depth revealed that abundance of the genus Clostridium (order Clostridiales) decreased in the presence of both S-NP (p < 0.01) and T-NP (p < 0.05). The abundance of the genus Vibrio (order Vibrionales) was likewise impacted in the presence of all the three NPs - S-NP (p < 0.01), T-NP (p < 0.05) and Z-NP (p < 0.05). Analyses at high taxon ranks such as phyla may not give a good representation of the nature of microbial community shifts, and at times may paint an erroneous picture. The use of full-length 16S rRNA gene sequences here yielded a greater taxonomic depth, and some shifts at the lower ranks were discernible.

Improving the stability of bacteriocin extracted from Enterococcus faecium by immobilization onto cellulose nanocrystals.

Enterococcus faecium (E. faecium) isolated from Vigna mungo (Black gram) produced bacteriocin that inhibits both Gram positive and Gram negative bacteria and better heat stability (100 °C for 30 min). The bacteriocin was sensitive to protease treatment and most active in acidic pH. Bacteriocin produced by Pediococcus acidilactici was used for comparison. To enhance stability for diversified applications, the bacteriocin was immobilized by physical adsorption onto cellulose nanocrystals (CNC) extracted from cotton linters. The bacteriocin immobilization yield was 64.91% for P. acidilactici and 53.63% for E. faecium. The bacteriocin immobilized CNC was characterized by DLS particle sizing, FTIR and AFM to evaluate size distribution, chemical nature and surface morphology. The bacteriocins immobilized on CNC showed 50% increase in stability in terms of antibacterial activity. The enzymatic synthesis of CNC in combination with physical adsorption immobilization method for bacteriocin makes it an efficient system of producing antibacterial nanofillers for food packaging and bio-composites applications.

Effect of Fenton's pretreatment on cotton cellulosic substrates to enhance its enzymatic hydrolysis response.

Fenton's reagent that generates reactive hydroxyl radical species was evaluated for its effectiveness as a pretreatment agent on cotton cellulosic substrates to increase its susceptibility to cellulase enzyme. Response surface methodology was used to optimize four different process variables viz., time of reaction; substrate size and concentrations of Fe2+ and H2O2. Overall, the cellulose substrates treated at 0.5 mM concentration of Fe2+, 2% concentration of H2O2 for a reaction period of 48 h gave the highest enzyme activity as determined using the response surface methodology. Cellulose substrates with high aspect ratio recorded better enzyme response than that with low aspect ratio which is supported by copper number estimation. The cellulosic substrate prepared using a combination of optimized Fenton's pretreatment conditions and/or enzyme hydrolysis were studied and characterized by atomic force microscopy and scanning electron microscopy. Additionally, degree of polymerization analysis gives further insight into the degradation during Fenton's reaction.

Silver-protein (core-shell) nanoparticle production using spent mushroom substrate.

A simple route for the synthesis of silver-protein (core-shell) nanoparticles using spent mushroom substrate (SMS) has been demonstrated in this work. SMS exhibits an organic surface that reduces silver ions and stabilizes the silver nanoparticles by a secreted protein. The silver nitrate solution incubated with SMS changed to a yellow color from 24 h onward, indicating the formation of silver nanoparticles. The purified solution yielded the maximum absorbance at 436 nm due to surface plasmon resonance of the silver nanoparticles. X-ray analysis of the freeze-dried powder of silver nanoparticles confirmed the formation of metallic silver. Transmission electron microscopic analysis of the samples showed a uniform distribution of nanoparticles, having an average size of 30.5 +/- 4.0 nm, and its corresponding electron diffraction pattern confirmed the face-centered cubic (fcc) crystalline structure of metallic silver. The characteristic fluorescence of the protein shell at 435 nm was observed for the silver nanoparticles in solution, when excited at 280 nm, while Fourier transform infrared (FTIR) spectroscopy confirmed the presence of a protein shell. The silver nanoparticles were found to be stable in solution for more than 6 months. It is observed that the reducing agents from the safflower stalks caused the reduction of silver ions while protein secreted by the fungus stabilized the silver nanoparticles. These silver nanoparticles showed excellent antibacterial activity against two representative bacteria, Staphylococcus aureus (Gram positive) and Klebsiella pneumoniae (Gram negative), in spite of the presence of an organic layer as a shell. Apart from ecofriendliness and easy availability, "SMS" as a biomanufacturing unit will give us an added advantage in ease of handling when compared to other classes of microorganisms.

Biomimetics of silver nanoparticles by white rot fungus, Phaenerochaete chrysosporium.

Extracellular synthesis of silver nanoparticles by a white rot fungus, Phaenerochaete chrysosporium is reported in this paper. Incubation of P. chrysosporium mycelium with silver nitrate solution produced silver nanoparticles in 24h. These silver nanoparticles were characterized by means of UV-vis spectroscopy, X-ray diffraction analysis, scanning electron microscopy, transmission electron microscopy, and photoluminescence spectroscopy. The synthesized silver nanoparticles absorbed maximum at 470 nm in the visible region. XRD spectrum of the silver nanoparticles confirmed the formation of metallic silver. The SEM characterization of the fungus reacted on the Ag+ indicated that the protein might be responsible for the stabilization of silver nanoparticles. This result was further supported by the TEM examination. Though shape variation was noticed, majority of the nanoparticles were found to be of pyramidal shape as seen under TEM. Photoluminescence spectrum showed a broad emission peak of silver nanoparticles at 423 nm when excited at 350 nm. Apart from eco-friendliness, fungus as bio-manufacturing unit will give us an added advantage in ease of handling when compared to other classes of microorganisms.

Spectroscopic characterization of zinc oxide nanorods synthesized by solid-state reaction.

Well-crystallized zinc oxide nanorods have been fabricated by single step solid-state reaction using zinc acetate and sodium hydroxide, at room temperature. The sodium lauryl sulfate (SLS) stabilized zinc oxide nanorods were characterized by using X-ray diffraction, Fourier transform infrared spectroscopy, transmission electron microscopy and photoluminescence spectroscopy. The X-ray diffraction revealed the wurtzite structure of zinc oxide. The size estimation by XRD and TEM confirmed that the ZnO nanorods are made of single crystals. The growth of zinc oxide crystals into rod shape was found to be closely related to its hexagonal nature. The mass ratio of SLS:ZnO in the nanorods was found to be 1:10 based on the thermogravimetric analysis. Blue shift of photoluminescence emission was noticed in the ZnO nanorods when compared to that of ZnO bulk. FT-IR analysis confirmed the binding of SLS with ZnO nanorods. Apart from ease of preparation, this method has the advantage of eco-friendliness since the solvent and other harmful chemicals were eliminated in the synthesis protocol.

Autofluorescence characterization of advanced glycation end products of hemoglobin.

This article describes the analysis of autofluorescence of advanced glycation end products of hemoglobin (Hb-AGE). Formed as a result of slow, spontaneous and non-enzymatic glycation reactions, Hb-AGE possesses a characteristic autofluorescence at 308/345 nm (lambda(ex)/lambda(em)). Even in the presence of heme as a quenching molecule, the surface presence of the glycated adduct gave rise to autofluorescence with the quantum yield of 0.19. The specificity of monoclonal antibody developed against common AGE structure with Hb-AGE was demonstrated using reduction in fluorescence polarization value due to increased molecular volume while binding. The formation of fluorescent adduct in hemoglobin in the advanced stage of glycation and the non-fluorescent HbA(1c) will be of major use in distinguishing and to know the past status of diabetes mellitus. While autofluorescence correlated highly with HbA(1c) value under in vivo condition (r = 0.85), it was moderate in the clinical samples (r = 0.55). The results suggest a non-linear relation between glycemia and glycation, indicating the application of Hb-AGE as a measure of susceptibility to glycation rather than glycation itself.