In this study, mycelial filtrate of Aspergillus terreus BA6 was used to reduce AgNO3 to form silver nanoparticles (AgNPs). The effect of seven independent variables on the diameter of AgNPs was studied by applying design of experiments (DOE). At optimal conditions, the diameter of AgNPs was reduced by approximately 26.7% compared to the basal culture condition and AgNO3 concentration was found to be the most significant factor affecting the diameter of AgNPs. A. terreus nano-Ag was characterized using UV-visible spectroscopy, transmission electron microscopy, energy dispersive X-ray (EDX), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and Zeta potential. The maximum UV absorption was obtained at 420 nm and the microscopic results showed particles with narrow size distribution ranging from 7 to 23 nm. XRD pattern of AgNPs revealed four diffraction peaks of metallic silver and the EDX spectrum showed a strong signal attributed to Ag nano-crystals. AgNPs mycofabricated by A. terreus showed potent minimum inhibitory concentration (MIC) and broad minimum bactericidal/fungicidal concentration (MBC/MFC) against 12 reference microorganisms. The MIC and MBC/MFC values of AgNPs were 0.312 to 1.25 μg/ml and 0.625 to 10 μg/ml, respectively. Nevertheless, AgNPs did not demonstrate any antagonistic activity against Coxsackie B virus. The in vitro cytotoxicity of the mycosynthesized AgNPs showed significant antitumor activity against adenocarcinoma epithelial cells from human breast cancer (Mcf-7) cell line with an inhibitory concentration (IC50) of 87.5 μg/ml.
Our previous study demonstrated that a new Bacillus subtilis AD35 was the first Bacillus sp. so far possessing the capability to produce the antibacterial compound di-(2-ethylhexyl) phthalate (DEHP) [Lotfy WA, Mostafa SW, Adel AA, et al. Production of DEHP by Bacillus subtilis AD35: isolation, purification, characterization and biological activities. Microb Pathog. 2018;124:89-100]. In this study, we investigate the process parameters influencing DEHP production by B. subtilis AD35. The best culture medium, carbon and nitrogen source that significantly affected DEHP synthesis were beef-yeast-peptone medium, starch and L-asparagine, respectively. A fractional factorial design (FFD) was applied to screen the fermentation conditions affecting DEHP formation. Agitation rate, inoculum size and sodium chloride concentration were found to be the major factors affecting the synthesis of DEHP. Sequentially, response surface methodology (RSM) was implemented and a polynomial model was developed to predict maximum DEHP activities against Escherichia coli, methicillin-resistant Staphylococcus aureus (MRSA), Salmonella typhimurium, Staphylococcus aureus, Staphylococcus epidermidis, Listeria monocytogenes, Pseudomonas aeruginosa and Aeromonas hydrophila. At optimized conditions, agitation rate, 220 rpm; inoculum size, 0.075% (v/v) and sodium chloride concentration, 0.75% (w/v), DEHP activity markedly increased to 24.7 AU, which represents an approximately 707% increase when compared to the pre-optimized conditions. The work reported here represents the first optimization of the process parameters influencing DEHP synthesis by B. subtilis AD35.
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