The search for biocompatible nanoparticles with vast applicability has impacted on exploration of various biomaterials for the synthesis of mono and bimetallic nanoparticles. Xylanase is widely regarded as an industrially important enzyme but its potentials in nanotechnological applications are yet to be fully explored. The current study investigates the exploit of xylanases of Aspergillus niger L3 (NE) and Trichoderma longibrachiatum L2 (TE) produced through valorization of corn‐cob, to synthesize silver‐gold alloy nanoparticles (Ag‐AuNPs). Characterization of the Ag‐AuNPs involved UV–vis spectroscopy, Fourier transform infrared spectroscopy (FTIR), and field emission scanning electron microscopy and transmission electron microscopy, while their prospective use as antimicrobial, antioxidant, catalytic, anticoagulant, and thrombolytic agents were studied. The biosynthesized Ag‐AuNPs were ruby red and light purple with surface plasmon resonance at 520 and 534 nm for NEAg‐AuNPs and TEAg‐AuNPs, respectively; while FTIR showed that protein molecules capped and stabilized the nanoparticles. The Ag‐AuNPs were anisotropic with spherical, oval, and irregular shapes having sizes ranging from 6.98 to 52.51 nm. The nanoparticles appreciably inhibited the growth of tested clinical bacteria (23.40–90.70%) and fungi (70.10–89.05%), and also scavenged 2,2‐diphenyl‐1‐picrylhydrazyl (48.51–53.79%) and hydrogen peroxide (80.5–95.50%). Furthermore, the Ag‐AuNPs degraded malachite green (91.39%) and methylene blue (47.10%). Moreover, the Ag‐AuNPs displayed outstanding anticoagulant and thrombolytic activities using human blood. This study further emphasizes the significance of xylanases in nanobiotechnology as it has established the potential of xylanases to synthesize Ag‐AuNPs, which is being reported for the first time.