Heavy metals are environmental pollutants affect the integrity and distribution of living organisms in the ecosystem and also humans across the food chain. The study targeted the removal of copper (Cu2+) from aqueous solutions, depending on the biosorption process. The bacterial candidate was identified using 16S rRNA sequencing and phylogenetic analysis, in addition to morphological and cultural properties as Azotobacter nigricans NEWG-1. The Box-Behnken design was applied to optimize copper removal by Azotobacter nigricans NEWG-1 and to study possible interactive effects between incubation periods, pH and initial CuSO4 concentration. The data obtained showed that the maximum copper removal percentage of 80.56% was reached at run no. 12, under the conditions of 200 mg/L CuSO4, 4 days' incubation period, pH, 8.5. Whereas, the lowest Cu2+ removal (12.12%) was obtained at run no.1. Cells of Azotobacter nigricans NEWG-1 before and after copper biosorption were analyzed using FTIR, EDS and SEM. FTIR analysis indicates that several functional groups have participated in the biosorption of metal ions including hydroxyl, methylene, carbonyl, carboxylate groups. Moreover, the immobilized bacterial cells in sodium alginate-beads removed 82.35 ± 2.81% of copper from the aqueous solution, containing an initial concentration of 200 mg/L after 6 h. Azotobacter nigricans NEWG-1 proved to be an efficient biosorbent in the elimination of copper ions from environmental effluents, with advantages of feasibility, reliability and eco-friendly.