This study investigated the green biosynthesis of gold (Au) and silver-gold alloy (Ag-Au) nanoparticles using cell-free extract of Bacillus safensis LAU 13 strain (GenBank accession No: KJ461434). The biosynthesized AuNPs and Ag-AuNPs were characterized using UV-Vis spectroscopy, Fourier-transform infrared spectroscopy, and transmission electron microscopy. Evaluation of the antifungal activities, degradation of malachite green, anti-coagulation of blood, and thrombolysis of human blood clot by the biosynthesized nanoparticles were investigated. The AuNPs and Ag-AuNPs had maximum absorbance at 561 and 545 nm, respectively. The FTIR peaks at 3318, 2378, 2114, 1998, 1636, 1287, 446, 421 cm for AuNPs; and 3310, 2345, 2203, 2033, 1636, 1273, 502, 453, 424 cm for Ag-AuNPs indicated that proteins were the capping and stabilization molecules in the biosynthesized nanoparticles. The particles were fairly spherical in shape with size of 10-45 nm for AuNPs and 13-80 nm for Ag-AuNPs. Moreover, energy dispersive X-ray analysis of AuNPs revealed gold as the most prominent metal in the AuNPs solution, while silver and gold were the most prominent in the case of Ag-AuNPs. Selected area electron diffraction showed the biosynthesized nanoparticles as crystal structures with ring shape pattern. AuNPs and Ag-AuNPs displayed growth inhibitions of 66.67-90.78% against strains of Aspergillus fumigatus and A. niger at concentration of 200 μg/ml , and remarkable degradation (> 90%) of malachite green after 48 h. Furthermore, the nanoparticles prevented coagulation of blood, and also completely dissolved blood clots, indicating the biomedical potential of AuNPs and Ag-AuNPs in the management of blood coagulation disorders. This is the first report of the synthesis of AuNPs and Ag-AuNPs using a strain of B. safensis for biomedical and catalytic applications.