RESUMO
The present study examined the biosynthesis and characterization of selenium nanoparticles (SeNPs) using two contrasting endophytic selenobacteria, one Gram-positive (Bacillus sp. E5 identified as Bacillus paranthracis) and one Gram-negative (Enterobacter sp. EC5.2 identified as Enterobacter ludwigi), 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 B. paranthracis and E-SeNPs from E. ludwigii) 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 B. paranthracis. 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.
RESUMO
The high P retention of acidic Andisols makes necessary to increase our technological approaches in pasture management in the animal system production. Here, we evaluated the clay- or nanoclay-acid phosphatase complexes for improving phosphorus mineralization from degraded cattle dung. We implemented an immobilization mechanism of acid phosphatase (AP) using natural clays (allophanic and montmorillonite) and nanoclays as support materials. Also, we evaluated the mineralization of organic P containing in decomposed cattle dung with clay- and nanoclay-AP complexes by incubation studies. Clays and nanoclays were characterized by microscopy techniques as atomic force and confocal-laser scanning microscopy. We found that these support materials stabilized AP by encapsulation. Our results showed that immobilization on allophanic or montmorillonite materials improved both the specific activity (4-48%) and the V(max) (28-38%) of AP. Moreover, the enzyme had a better performance when immobilized on clay and nanoclay from Andisol than on montmorillonite materials. Phosphorous mineralization of cattle dung was regulated by water-soluble P present in the dung and P re-adsorption on allophanic materials. However, we were able to detect a potential capacity of AP immobilized on allophanic nanoclays as the best alternative for P mineralization. Further research with initially low water-soluble P containing organic materials is required to quantify the P mineralization potential and bioavailability of P from dung.