The current investigation addressed the green synthesis of silver nanoparticles (AgNPs) using newly isolated silver-resistant rare actinomycetes, Glutamicibacter nicotianae SNPRA1 and Leucobacter aridicollis SNPRA2, and investigated their impact on the mycotoxigenic fungi Aspergillus flavus ATCC 11498 and Aspergillus ochraceus ATCC 60532. The formation of AgNPs was evidenced by the reaction’s color change to brownish and the appearance of the characteristic surface plasmon resonance. The transmission electron microscopy of biogenic AgNPs produced by G. nicotianae SNPRA1 and L. aridicollis SNPRA2 (designated Gn-AgNPs and La-AgNPs, respectively) revealed the generation of monodispersed spherical nanoparticles with average sizes of 8.48 ± 1.72 nm and 9.67 ± 2.64 nm, respectively. Furthermore, the XRD patterns reflected their crystallinity and the FTIR spectra demonstrated the presence of proteins as capping agents. Both bioinspired AgNPs exhibited a remarkable inhibitory effect on the conidial germination of the investigated mycotoxigenic fungi. The bioinspired AgNPs caused an increase in DNA and protein leakage, suggesting the disruption of membrane permeability and integrity. Interestingly, the biogenic AgNPs completely inhibited the production of total aflatoxins and ochratoxin A at concentrations less than 8 μg/mL. At the same time, cytotoxicity investigations revealed the low toxicity of the biogenic AgNPs against the human skin fibroblast (HSF) cell line. Both biogenic AgNPs exhibited feasible biocompatibility with HSF cells at concentrations up to 10 μg/mL and their IC50 values were 31.78 and 25.83 μg/mL for Gn-AgNPs and La-AgNPs, respectively. The present work sheds light on the antifungal prospect of the biogenic AgNPs produced by rare actinomycetes against mycotoxigenic fungi as promising candidates to combat mycotoxin formation in food chains at nontoxic doses.
The biosynthesis of nanoparticles using green technology is emerging as a cost-efficient, eco-friendly and risk-free strategy in nanotechnology. Recently, tellurium nanoparticles (TeNPs) have attracted growing attention due to their unique properties in biomedicine, electronics, and other industrial applications. The current investigation addresses the green synthesis of TeNPs using a newly isolated mangrove-associated bacterium, Gayadomonas sp. TNPM15, and their impact on the phytopathogenic fungi Fusarium oxysporum and Alternaria alternata. The biogenic TeNPs were characterized using transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), Raman spectroscopy and Fourier transform infrared (FTIR). The results of TEM revealed the intracellular biosynthesis of rod-shaped nanostructures with a diameter range from 15 to 23 nm and different lengths reaching up to 243 nm. Furthermore, the successful formation of tellurium nanorods was verified by SEM-EDX, and the XRD pattern revealed their crystallinity. In addition, the FTIR spectrum provided evidence for the presence of proteinaceous capping agents. The bioinspired TeNPs exhibited obvious inhibitory effect on the spores of both investigated phytopathogens accomplished with prominent ultrastructure alternations, as evidenced by TEM observations. The biogenic TeNPs impeded spore germination of F. oxysporum and A. alternata completely at 48.1 and 27.6 µg/mL, respectively. Furthermore, an increase in DNA and protein leakage was observed upon exposure of fungal spores to the biogenic TeNPs, indicating the disruption of membrane permeability and integrity. Besides their potent influence on fungal spores, the biogenic TeNPs demonstrated remarkable inhibitory effects on the production of various plant cell wall-degrading enzymes. Moreover, the cytotoxicity investigations revealed the biocompatibility of the as-prepared biogenic TeNPs and their low toxicity against the human skin fibroblast (HSF) cell line. The biogenic TeNPs showed no significant cytotoxic effect towards HSF cells at concentrations up to 80 μg/mL, with a half-maximal inhibitory concentration (IC50) value of 125 μg/mL. The present work spotlights the antifungal potential of the biogenic TeNPs produced by marine bacterium against phytopathogenic fungi as a promising candidate to combat fungal infections.
The main challenge of astaxanthin extraction is to provide an eco-friendly method of extraction instead of chemical methods that harm human health. This study provided an eco-friendly method for astaxanthin extraction using two bacterial and fungal probiotics (Bifidobacterium lactis, Lactobacillus lactis, Candida utilis, and Saccharomyces cerevisiae, respectively) and determined the astaxanthin concentration by high-performance liquid chromatography (HPLC) analysis. The results showed that the highest concentration was obtained by S. cerevisiae (45.69 µg/g). Several biological tests were done on the exoskeleton containing astaxanthin of crawfish. Antifungal activity was effective against C. utilis (inhibition zone is 12.3 ± 0.5 mm). The scavenging percentage of 2,2-diphenyl-1-picrylhydrazyl (DPPH scavenging percentage) was 72.1% at 1000 µg/mL concentration of exoskeleton containing astaxanthin. The Hemolysis inhibition percentage was 65% at the same concentration used previously. Furthermore, the IC50 value of human liver cancer cell line (HepG2), human hepatocellular carcinoma (HCT), and breast cancer cell line MCF-7 were 24 µg/mL, 11 µg/mL, and 9.5 µg/mL, respectively. The least cell viability percentage was 19% (using breast cancer cell line (MCF-7)) at 100 µg/mL of exoskeleton containing astaxanthin. Thus, using microorganisms can be an alternative and promising way of astaxanthin extraction. Furthermore, purification of extracted astaxanthin is essential for medical applications.
D-Tagatose is a low-calorie sugar substitute that has gained increased attention as a functional sweetener owing to its nutraceutical and prebiotic properties. Traditionally, D-tagatose is produced via the enzymatic conversion of L-galactose to D-tagatose by L-arabinose isomerase (L-AI). Nonetheless, the most reported L-AI enzymes are ion-dependent enzymes requiring Mn2+ and/or Co2+ as cofactors for their reactions, which limits their application due to safety and health concerns. Herein, we addressed the facile bioconversion of L-galactose to D-tagatose using a novel recombinant metallic-ions-independent L-AI derived from endophytic Bacillus amyloliquefaciens CAAI isolated from cantaloupe fruits. The ORF (1500 bp) of the L-arabinose isomerase gene (araA) was cloned and over-expressed in Escherichia coli. The recombinant enzyme (BAAI) was purified to homogeneity using Ni-NTA affinity chromatography, yielding a single distinct band with an apparent molecular mass of approximately 59 kDa as deduced from SDS-PAGE analysis. The purified enzyme showed optimum activity at pH and temperature of 7.5 and 45 °C, respectively, with obvious enzymatic activity in the presence of ethylenediaminetetraacetic acid (EDTA), indicating the metallic-ions independence from BAAI. The Km values of BAAI for D-galactose and L-arabinose were 251.6 mM and 92.8 mM, respectively. The catalytic efficiency (kcat/Km) values for D-galactose and L-arabinose were found to be 2.34 and 46.85 mM–1 min–1, respectively. The results revealed the production of 47.2 g/L D-tagatose from D-galactose (100 g/L) with 47.2% bioconversion efficiency in a metallic-ions-free reaction system that could be implemented in safe-production of food-grade low-calorie sweetener, D-tagatose.
Schistosomiasis is a severe illness carried by snails of the genus Biomphalaria, which have a large distribution in Egypt. Control of the intermediate host snail is critical in limiting schistosomiasis spread. On the topic of snails’ management, bioactive products extracted from aquatic invertebrates have gained more interest. Here, for the first time, the current research intended to identify the bioactive compounds present in hemolymph collected from the freshwater crayfish Procambarus clarkii and to assess its molluscidal activity against Biomphalaria alexandrina snails. Results showed that following exposure to the sub-lethal concentrations (LC10 34.9 and LC25 42.8 mg/l) of hemolymph, a reduction in survival and reproductive rates of B. alexandrina was detected, as well as a substantial increase in transaminases (AST and ALT) activities, while a reduction in total protein and albumin levels. Also, levels of testosterone and 17β-estradiol hormones decreased compared to the control group. Histopathological examination of the digestive gland of treated B. alexandrina displayed a shrinkage of the tubules and an increase in the inter-tubular spaces, and also, degeneration in the oocytes and spermatocytes of the hermaphrodite glands with a loss of connective tissues between the acini was observed. Thus, based on these results, a new strategy should be developed for a better use of these agents in schistosomiasis control.
Controlling Biomphalaria species with plant products is crucial in the treatment of schistosomiasis since it is eco-friendly and generally less harmful to non-target organisms. The goal of this study was to determine the molluscicidal activity of an aqueous extract of Arabidopsis thaliana leaves against Biomphalaria glabrata. A preliminary phytochemical screening test revealed the presence of flavonoids, terpenoids, steroids, anthraquinones, alkaloids, saponins, tannins, and carbohydrates. Following exposure to sub-lethal concentrations (LC10 75.62 mg/L and LC25 90.52 mg/L) of aqueous extract of A. thaliana, a reduction in survival, reproductive, and fecundity rates of B. glabrata were detected, as well as a substantial decrease in GSH, CAT, and SOD, while increasing MDA and NO levels. In addition, there was an increase in liver and renal functions as well as lipid profiles, compared to the control group. Histopathological examination of the digestive gland of treated B. glabrata showed a shrinkage of the tubules and an increase in the inter-tubular spaces as well as degeneration in the oocytes and spermatocytes of the hermaphrodite glands with a loss of connective tissues between the acini. Also, the comet assay revealed a genotoxic effect of aqueous extract of A. thaliana on B. glabrata, with a significant increase in the tail moment, tail length, and DNA percentage reflecting DNA damage compared to the control group. Conclusively, Arabidopsis thaliana aqueous extract acts as a natural molluscicidal agent against Biomphalaria glabrata.