Production, purification, and characterization of p-diphenol oxidase (PDO) enzyme from lignolytic fungal isolate Schizophyllum commune MF-O5.

Fungi are producers of lignolytic extracellular enzymes which are used in industries like textile, detergents, biorefineries, and paper pulping. This study assessed for the production, purification, and characterization of novel p-diphenol oxidase (PDO; laccase) enzyme from lignolytic white-rot fungal isolate. Fungi samples collected from different areas of Pakistan were initially screened using guaiacol plate method. The maximum PDO producing fungal isolate was identified on the basis of ITS (internal transcribed spacer sequence of DNA of ribosomal RNA) sequencing. To get optimum enzyme yield, various growth and fermentation conditions were optimized. Later PDO was purified using ammonium sulfate precipitation, size exclusion, and anion exchange chromatography and characterized. It was observed that the maximum PDO producing fungal isolate was Schizophyllum commune (MF-O5). Characterization results showed that the purified PDO was a monomeric protein with a molecular mass of 68 kDa and showed stability at lower temperature (30 °C) for 1 h. The Km and Vmax values of the purified PDO recorded were 2.48 mM and 6.20 U/min. Thermal stability results showed that at 30 °C PDO had 119.17 kJ/K/mol Ea value and 33.64 min half-life. The PDO activity was stimulated by Cu2+ ion at 1.0 mM showing enhanced activity up to 111.04%. Strong inhibition effect was noted for Fe2+ ions at 1 mM showing 12.04% activity. The enzyme showed stability against 10 mM concentration oxidizing reducing agents like DMSO, EDTA, H2O2, NaOCl, and urea and retained more than 75% of relative activity. The characterization of purified PDO enzyme confirmed its tolerance against salt, metal ions, organic solvents, and surfactants indicating its ability to be used in the versatile commercial applications.

Synthesis of Biogenic Silver Nanocatalyst and their Antibacterial and Organic Pollutants Reduction Ability.

Plant-mediated nanoparticles are gaining popularity due to biologically active secondary metabolites that aid in green synthesis. This study describes a simple, environmentally friendly, dependable, and cost-effective production of silver nanoparticles utilizing Cucumis sativus and Aloe vera aqueous leaf extracts. The aqueous leaf extracts of Cucumis sativus and Aloe vera, which worked as a reducing and capping agent, were used to biosynthesize silver nanoparticles (AgNPs). The formation of surface plasmon resonance peaks at 403 and 405 nm corresponds to the formation of colloidal Ag nanoparticles. Similarly, the Bragg reflection peaks in X-ray diffraction patterns observed at 2θ values of 38.01°, 43.98°, 64.24°, and 77.12° representing the planes of [111], [200], [220], and [311] correspond to the face-centered cubic crystal structure of silver nanoparticles. Fourier transform infrared spectroscopy confirms that bioactive chemicals are responsible for the capping of biogenic silver nanoparticles. The size, structure, and morphology of AgNPs with diameters ranging from 8 to 15 nm were examined using transmission electron microscopy. Water contamination by azo dyes and nitrophenols is becoming a more significant threat every day. The catalytic breakdown of organic azo dye methyl orange (MO) and the conversion of para-nitrophenol (PNP) into para-aminophenol using sodium borohydride was evaluated using the prepared biogenic nanoparticles. Our nanoparticles showed excellent reduction ability against PNP and MO with rate constants of 1.51 × 10-3 and 6.03 × 10-4s-1, respectively. The antibacterial activity of the nanomaterials was also tested against four bacteria: Staphylococcus aureus, Klebsiella pneumoniae, Enterobacter, and Streptococcus pneumoniae. These biogenic AgNPs displayed effective catalytic and antibacterial characteristics by reducing MO and PNP and decreasing bacterial growth.

Use of scanning electron microscopy to analyze sculpturing pattern and internal features of pollen grain wall in some members of Astragaleae (subfamily: Papilionoidae).

Palynological features of tribe Astagaleae L. have been observed with scanning electron microscopy (SEM), to assess features that can be used to re-examine the placement and taxonomic position of tribe. The palynomorph attributes of 10 species included in three genera of Astragaleae (Astragalus L., Glycyrrhiza L., and Oxytropis DC.) are investigated. The assignment of genus Oxytropis in tribe Astragaleae was also reevaluated based on pollen characters. Pollen appear to be prolate, prolate-spheroidal, and subprolate. Polar axis size (P) varies from 31.3 µm ±1 to 17.5 µm ±1.4 and equatorial axis size (E) varies from 22.2 µm ±1.8 to 13.1 µm ±0.9. Prominent apertures found consistently dispersed along the surface of pollen. Three kinds of ornamentation pattern were recorded, that is, reticulate or reticulate-perforate and perforate. The studied species display variation in equatorial and polar diameter, aperturation and sculpturing pattern, exine thickness, and pollen shape. On the basis of descriptive pollen features, a dichotomous taxonomic key and UPGMA analysis has been made for effortless and quick identification. The study concluded that disparities of the entire features are typically unrelenting and concerned with the species and shows potential systematic significance. The combination of palynological attributes in association with additional traits has prospective for systematic identification at species and genus level.

Exceptional antibacterial and cytotoxic potency of monodisperse greener AgNPs prepared under optimized pH and temperature.

In the present work, silver nanoparticles were prepared by using the extract of Camellia Sinensis. The extract contains phytochemicals which are mainly polyphenols acting as the natural reducing and stabilizing agents leading to the formation of uniformly dispersed and stabilized silver nanoparticles. The synthesis of silver nanoparticles was significantly influenced by the impact of the pH, as well as temperature conditions. It was found that at pH 5 and 25 °C, nanoparticles of different morphologies (spherical, polygonal, capsule) and sizes were formed. However, with the increase in temperature from 25 °C to 65 °C but at the same pH, these particles started attaining the spherical shape of different sizes owing to an increase in the reduction rate. Furthermore, for the reaction of the mixture at 65 °C, an increase in pH from 5 to 11 led to an increase in the monodispersity of spherically shaped nanoparticles, attributed to the hydroxide ions facilitated reduction. The prepared nanoparticles were investigated for their antibacterial activity using Nathan's Agar Well-Diffusion method. It was found that AgNPs prepared at pH 9 and 65 °C demonstrated strong antibacterial activity against gram-negative Escherichia coli in contrast to gram-positive Staphylococcus aureus. In reference to the cytotoxic potency, the prepared AgNPs showed clear cytotoxicity for HeLa cells and showcased a close relationship between activity and concentration as evidenced by the decrease in the percentage (100 to 30%) of metabolically active cells up to 25 µM-75 µM concentration of silver nanoparticles.