Sans-Serramitjana Eulàlia, Gallardo-Benavente Carla, Melo Francisco, Pérez-Donoso José M, Rumpel Cornelia, Barra Patricio Javier, Durán Paola, Mora María de La Luz
Center of Plant, Soil Interaction and Natural Resources Biotechnology, Scientific and Biotechnological Bioresource Nucleus (BIOREN-UFRO), Universidad de La Frontera, Avenida Francisco Salazar 01145, P.O. Box 54-D, Temuco 4811230, Chile.
Centro Biotecnológico de Estudios Microbianos (CEBEM), Universidad de La Frontera, Temuco 4811230, Chile.
Microorganisms. 2023 Jun 16;11(6):1600. doi: 10.3390/microorganisms11061600.
The present study examined the biosynthesis and characterization of selenium nanoparticles (SeNPs) using two contrasting endophytic selenobacteria, one Gram-positive ( sp. E5 identified as ) and one Gram-negative ( sp. EC5.2 identified as ), for further use as biofortifying agents and/or for other biotechnological purposes. We demonstrated that, upon regulating culture conditions and selenite exposure time, both strains were suitable "cell factories" for producing SeNPs (B-SeNPs from and E-SeNPs from ) with different properties. Briefly, dynamic light scattering (DLS), transmission electron microscopy (TEM), and atomic force microscopy (AFM) studies revealed that intracellular E-SeNPs (56.23 ± 4.85 nm) were smaller in diameter than B-SeNPs (83.44 ± 2.90 nm) and that both formulations were located in the surrounding medium or bound to the cell wall. AFM images indicated the absence of relevant variations in bacterial volume and shape and revealed the existence of layers of peptidoglycan surrounding the bacterial cell wall under the conditions of biosynthesis, particularly in the case of . Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR), energy-dispersive X-ray (EDS), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS) showed that SeNPs were surrounded by the proteins, lipids, and polysaccharides of bacterial cells and that the numbers of the functional groups present in B-SeNPs were higher than in E-SeNPs. Thus, considering that these findings support the suitability of these two endophytic stains as potential biocatalysts to produce high-quality Se-based nanoparticles, our future efforts must be focused on the evaluation of their bioactivity, as well as on the determination of how the different features of each SeNP modulate their biological action and their stability.
本研究使用两种截然不同的内生硒细菌(一种革兰氏阳性菌(鉴定为sp. E5)和一种革兰氏阴性菌(鉴定为sp. EC5.2))研究了硒纳米颗粒(SeNPs)的生物合成及特性,以便进一步用作生物强化剂和/或用于其他生物技术目的。我们证明,通过调节培养条件和亚硒酸盐暴露时间,这两种菌株都是生产具有不同特性的SeNPs(来自的B - SeNPs和来自的E - SeNPs)的合适“细胞工厂”。简而言之,动态光散射(DLS)、透射电子显微镜(TEM)和原子力显微镜(AFM)研究表明,细胞内的E - SeNPs(56.23±4.85 nm)直径小于B - SeNPs(83.44±2.90 nm),并且两种制剂都位于周围介质中或与细胞壁结合。AFM图像表明细菌体积和形状没有相关变化,并揭示了在生物合成条件下细菌细胞壁周围存在肽聚糖层,特别是在的情况下。拉曼光谱、傅里叶变换红外光谱(FTIR)、能量色散X射线(EDS)、X射线衍射(XRD)和X射线光电子能谱(XPS)表明,SeNPs被细菌细胞的蛋白质、脂质和多糖包围,并且B - SeNPs中存在的官能团数量高于E - SeNPs。因此,鉴于这些发现支持这两种内生菌株作为生产高质量硒基纳米颗粒的潜在生物催化剂的适用性,我们未来的工作必须集中在评估它们的生物活性,以及确定每个SeNP的不同特征如何调节它们的生物作用及其稳定性上。