In our present study, we investigated the mechanism of Cd(II) biosorption from aqueous solution by Pseudomonas plecoglossicida using different instrumental techniques. The adsorption kinetics fitted well with the pseudo second-order model, suggesting that the Cd(II) adsorption by P. plecoglossicida consisted of a chemisorption and a physisorption process. Compared with the dead P. plecoglossicida cells, the live cells demonstrated the same adsorption capacity of Cd(II). Scanning electron microscope with energy dispersive X-ray spectroscopy analysis revealed that the main mechanism of adsorption was the combination of Cd(II) with the organic functional groups in the cell wall of P. plecoglossicida. Furthermore, Fourier transform infrared spectroscopic analysis of the metal-loaded biosorbent confirmed the participation of -NH, -OH, -CH, and -CONH groups in the uptake of Cd(II). Moreover, cation transport test revealed that ionic exchange interactions were involved in the Cd(II) adsorption. However, it only played a minor role in the Cd(II) biosorption process.