Botany Department, Faculty of Science, Tanta University, Tanta, Egypt.
Biochemistry Department, Faculty of Science, Damanhour University, Damanhour, Egypt.
Microb Cell Fact. 2023 Nov 6;22(1):228. doi: 10.1186/s12934-023-02238-4.
Mycosynthesis of silver nanoparticles (SNPs) offers a safe, eco-friendly, and promising alternative technique for large-scale manufacturing. Our study might be the first report that uses mycelial filtrate of an endophytic fungus, Aspergillus flavipes, for SNPs production under optimal conditions as an antimicrobial agent against clinical multidrug-resistant (MDR) wound pathogens.
In the present study, among four different endophytic fungi isolated from leaves of Lycium shawii, the only one isolate that has the ability to mycosynthesize SNPs has been identified for the first time as Aspergillus flavipes AUMC 15772 and deposited in Genebank under the accession number OP521771. One variable at a time (OVAT) and Plackett Burman design (PBD) were conducted for enhancing the production of mycosynthesized SNPs (Myco-SNPs) through optimization using five independent variables. The overall optimal variables for increasing the mycosynthesis of SNPs from mycelial filtrate of A. flavipes as a novel endophytic fungus were a silver nitrate concentration of 2 mM, a pH of 7.0, an incubation time of 5 days, and a mycelial filtrate concentration of 30% in dark conditions. UV-visible spectroscopy (UV-Vis), Fourier transform infrared spectroscopy (FT-IR), X-ray spectroscopy (XRD), Transmission electron microscopy (TEM), and Selected-Area Electron Diffraction (SAED) patterns were used to characterize Myco-SNPs, which showed the peak of absorbance at 420 nm, and FTIR showed the bands at 3426.44, 2923.30, 1681.85, 1552.64, and 1023.02 cm-1, respectively, which illustrated the presence of polyphenols, hydroxyl, alkene, nitro compounds, and aliphatic amines, respectively. The XRD pattern revealed the formation of Myco-SNPs with good crystal quality at 2θ = 34.23° and 38.18°. The TEM image and SAED pattern show the spherical crystalline shape of Myco-SNPs with an average size of 6.9232 nm. High antibacterial activity of Myco-SNPs was recorded against MDR wound pathogens as studied by minimum inhibitory concentrations ranging from 8 to 32 µg/mL, time kill kinetics, and post-agent effects. Also, in vitro cell tests indicated that Myco-SNPs support the cell viability of human skin fibroblast cells as a nontoxic compound.
The obtained results revealed the successful production of Myco-SNPs using the mycelial filtrate of A. flavipes, which may be a promising nontoxic alternative candidate for combating MDR wound pathogens.
银纳米粒子(SNP)的真菌合成提供了一种安全、环保且有前途的大规模制造技术。我们的研究可能是首次使用内生真菌黄曲霉的菌丝滤液在最佳条件下生产 SNP 作为抗临床多药耐药(MDR)伤口病原体的抗菌剂。
在本研究中,从枸杞叶中分离出的四种不同内生真菌中,只有一种能够真菌合成 SNP 的真菌被首次鉴定为黄曲霉 AUMC 15772,并在 Genebank 中以 OP521771 登录号进行了存储。使用五个独立变量进行一次变量(OVAT)和 Plackett Burman 设计(PBD),以优化通过优化从黄曲霉菌丝滤液中合成 SNP(Myco-SNP)的产量。增加新型内生真菌黄曲霉菌丝滤液中 SNP 真菌合成的总体最佳变量是硝酸银浓度为 2mM、pH 值为 7.0、培养时间为 5 天和黑暗条件下菌丝滤液浓度为 30%。使用紫外可见分光光度法(UV-Vis)、傅里叶变换红外光谱(FT-IR)、X 射线光谱(XRD)、透射电子显微镜(TEM)和选区电子衍射(SAED)图谱对 Myco-SNP 进行了表征,其在 420nm 处显示出最大吸收峰,FTIR 显示出分别在 3426.44、2923.30、1681.85、1552.64 和 1023.02cm-1 处的峰,分别表示多酚、羟基、烯烃、硝基化合物和脂肪胺的存在。XRD 图谱表明,在 2θ=34.23°和 38.18°处形成了具有良好结晶质量的 Myco-SNP。TEM 图像和 SAED 图谱显示 Myco-SNP 的球形结晶形状,平均尺寸为 6.9232nm。通过最低抑菌浓度范围为 8 至 32μg/mL、时间杀伤动力学和后剂效应研究,记录了 Myco-SNP 对 MDR 伤口病原体的高抗菌活性。此外,体外细胞试验表明,Myco-SNPs 支持人皮肤成纤维细胞的细胞活力,是一种无毒化合物。
使用黄曲霉的菌丝滤液成功生产了 Myco-SNP,这可能是一种有前途的无毒替代 MDR 伤口病原体的候选物。