Optimizing mycelial protein yield in Pleurotus djamor via ARTP mutagenesis and hybridization strategies.

With the world's population rapidly increasing, the demand for high-quality protein is on the rise. Edible fungi breeding technology stands as a crucial avenue to obtain strains with high yield, high-quality protein, and robust stress resistance. To address the protein supply gap, Atmospheric and Room Temperature Plasma (ARTP) mutagenesis, and spore hybridization techniques were employed to enhance Pleurotus djamor mycelium protein production. Beginning with the original strain Pleurotus djamor JD-1, ARTP was utilized to mutate spore suspension. The optimal treatment time for Pleurotus djamor spores, determined to achieve optimal mortality, was 240 s. Through primary and secondary screenings, 6 mutant strains out of 39 were selected, exhibiting improved protein yield and growth rates compared to the original strain. Among these mutagenic strains, 240S-4 showcased the highest performance, with a mycelial growth rate of 9.5±0.71 mm/d, a biomass of 21.45±0.54 g/L, a protein content of 28.75±0.92%, and a remarkable protein promotion rate of 128.03±7.29%. Additionally, employing spore hybridization and breeding, 7 single-nuclei strains were selected for pin-two hybridization, resulting in 21 hybrid strains. The biomass and protein content of 9 hybrid strains surpassed those of the original strains. One hybrid strain, H-5, exhibited remarkable mycelial protein production, boasting a mycelial growth rate of 26.5±0.7 mm/d, a biomass of 21.70±0.46 g/L, a protein content of 28.44±0.22%, and a protein promotion rate of 128.02±1.73%. Notably, both strains demonstrated about a 28% higher mycelial protein yield than the original strains, indicating comparable effectiveness between hybrid breeding and mutagenesis breeding. Finally, we analyzed the original and selected strains by molecular biological identification, which further proved the effectiveness of the breeding method. These findings present novel insights and serve as a reference for enhancing edible fungi breeding, offering promising avenues to meet the escalating protein demand.

Engineering Bacterial Biofilm Development and Structure via Regulation of Silver Nanoparticle Density in Graphene Oxide Composite Coating.

Graphene-based composites have shown significant potential in the treatment of biofilm infections in clinical settings due to their exceptional antimicrobial properties and specific mechanisms. Nevertheless, a comprehensive understanding of the influence exerted by nanoparticles embedded in the composites on the development and structure of biofilms is still lacking. Here, we fabricate different graphene oxide-silver nanoparticle (GAg) composite-modified substrates (GAgS) with varying densities of silver nanoparticles (AgNPs) and investigate their effects on planktonic bacterial adhesion, subsequent biofilm formation, and mature biofilm structure. Our findings indicate that the initial attachment of Pseudomonas aeruginosa cells during biofilm formation is determined by the density of AgNPs on the GAgS surface. In contrast, the subsequent transition from adherent bacteria to the biofilm is determined by GAgS's synergistic antimicrobial effect. There exists a threshold for the inhibitory performance of GAgS, where the 20 µg/cm2 GAg composite completely prevents biofilm formation; below this concentration, GAgS delays the development of the biofilm and causes structural changes in the mature biofilm with enhanced bacterial growth and increased production of extracellular polymeric substance. More importantly, GAgS have minimal impact on mammalian cell morphology and proliferation while not inducing hemolysis in red blood cells. These results suggest that GAg composites hold promise as a therapeutic approach for addressing medical devices and implant-associated biofilm infections.

Polyethylene glycol modified graphene oxide-silver nanoparticles nanocomposite as a novel antibacterial material with high stability and activity.

Inorganic antibacterial nanomaterials play an increasingly important role in addressing the growing threat of drug-resistant bacteria. Graphene oxide-silver nanoparticles composite (GO-AgNPs), as a kind of inorganic nanomaterials, have excellent antibacterial properties, showing promising potential in biomedical field. However, GO-AgNPs are terribly prone to sedimentation due to aggregation in physiological solutions, along with its non-environmental issues during the synthesis process, seriously limits the antibacterial application of GO-AgNPs in the biomedical field. To solve this problem, herein, polyethylene glycol-graphene oxide-silver nanoparticles composite (GO-AgNPs-PEG) were prepared by modifying GO-AgNPs with polyethylene glycol to enhance their dispersion stability in physiological solutions. In addition, GO-AgNPs-PEG were prepared with using the natural product gallic acid as a reductant and stabilizer, exhibiting the characteristic of environmentally friendly. Meanwhile, the dispersion stability and antibacterial activity of GO-AgNPs-PEG were characterized by various technical methods, it was found that GO-AgNPs-PEG can be stably dispersed in a variety of physiological solutions (e.g., physiological saline, phosphate buffer solution, Luria-Bertani medium, Murashige and Skoog medium) for more than one week. Moreover, the antibacterial properties of GO-AgNPs-PEG in physiological solutions were significantly better than those of GO-AgNPs. Furthermore, it was discovered that the antibacterial mechanism of GO-AgNPs-PEG was probably associated to destroying the integrity of bacterial cell walls and membranes. The findings in this work can provide new ideas and references for the development of new inorganic antibacterial nanomaterials with stable dispersion in physiological solutions.

In Situ Green Synthesis of Graphene Oxide-Silver Nanoparticles Composite with Using Gallic Acid.

The adoption of plant-derived natural products to synthesize metal nanoparticles and their complexes has the advantages of mild reaction conditions, environmental protection, sustainability and simple operation compared with traditional physical or chemical synthesis methods. Herein, silver nanoparticles (AgNPs) were in situ synthesized on the surface of graphene oxide (GO) by a "one-pot reaction" to prepare graphene oxide-silver nanoparticles composite (GO-AgNPs) based on using AgNO3 as the precursor of AgNPs and gallic acid (GA) as the reducing agent and stabilizer. The size and morphology of GO-AgNPs were characterized by ultraviolet-visible spectrophotometer (Uv-vis), Fourier transform infrared spectroscopy (FT-IR), transmission electron microscope (TEM), X-ray diffractometer (XRD) and dynamic light scattering (DLS). The effects of pH, temperature, time and material ratio on the synthesis of GO-AgNPs were investigated experimentally. The results showed that ideal GO-AgNPs could be prepared under the conditions of pH = 9, 45°C, 2 h and the 2:1 of molar ratio of AgNO3 to GA. The AgNPs within GO-AgNPs are highly crystalline spherical particles with moderate density on the surface of GO, and the size of AgNPs is relatively uniform and determined to be about 8.19 ± 4.21 nm. The research results will provide new ideas and references for the green synthesis of metal nanoparticles and their complexes using plant-derived natural products as the reducing agent and stabilizer.

Functionalization and Antibacterial Applications of Cellulose-Based Composite Hydrogels.

Pathogens, especially drug-resistant pathogens caused by the abuse of antibiotics, have become a major threat to human health and public health safety. The exploitation and application of new antibacterial agents is extremely urgent. As a natural biopolymer, cellulose has recently attracted much attention due to its excellent hydrophilicity, economy, biocompatibility, and biodegradability. In particular, the preparation of cellulose-based hydrogels with excellent structure and properties from cellulose and its derivatives has received increasing attention thanks to the existence of abundant hydrophilic functional groups (such as hydroxyl, carboxy, and aldehyde groups) within cellulose and its derivatives. The cellulose-based hydrogels have broad application prospects in antibacterial-related biomedical fields. The latest advances of preparation and antibacterial application of cellulose-based hydrogels has been reviewed, with a focus on the antibacterial applications of composite hydrogels formed from cellulose and metal nanoparticles; metal oxide nanoparticles; antibiotics; polymers; and plant extracts. In addition, the antibacterial mechanism and antibacterial characteristics of different cellulose-based antibacterial hydrogels were also summarized. Furthermore, the prospects and challenges of cellulose-based antibacterial hydrogels in biomedical applications were also discussed.