Use of natural wool fiber supports in the fabrication of novel composite materials incorporating metal nanoparticles, which offer the possibility of “environmentally friendly” catalytic materials, has been investigated. The catalytic hydrogenation of cyclohexene to cyclohexane by palladium nanoparticles immobilized on wool (Pd/wool) was studied using moderate pressure of pure hydrogen gas. The performance of wool-supported catalysts was explored over a palladium nanoparticle loading ranging from 1.6 to 2.6 wt %. The effect of the catalytic testing conditions, including stirring rate, amount of reactants, gas pressure, and target temperature were explored. A systematic series of catalytic-activity tests carried out at 400 psi H2 for 5 and 24 h reaction times at 40 °C using a stirring rate 750 rpm allowed us to identify differences in performance within the series of Pd/wool nanocatalysts studied. The most catalytically active samples contained Pd nanoparticles with average sizes of ca. 5 nm located predominantly on the surface and within the topmost layer of wool fibers, making them more accessible to the reactants.