In the current study, new biomasses were prepared from fungi isolates including, Penicillium chrysogenum and Aspergillus ustus and utilized for the removal of heavy metals. During the study, the influences of various experimental factors including solution pH, temperature, and contact time on biosorption process were investigated. The synthesized fungi biosorbents were successfully applied for the removal of heavy metals from various waste effluents. The experimental results were indicated the excellent uptake property of the materials for target heavy metals. The average concentrations of Cd (II), Cu (II), and Pb (II) in different wastewater samples collected from Western, Eastern, and Northern region of Saudi Arabia were found to be in the range of 2.6–234 µg/L. The concentrations of Cu(II) in all the analyzed contaminated water samples were fairly exceeded the maximum concentration level for drinking water stipulated by World Health Organization. The applied biosorption technique for heavy metal removal is new, low cost and was adopted in eco-friendly way.