Background: Toxic heavy metals, such as lead, are widely used in industry and may cause serious health problems and ecological hazards for living organisms. Objectives: The current study aimed to investigate the removal efficiency of lead by Lactobacillus strains using a methodological approach. Methods: After selecting the bacteria with the maximum metals removal ability, experiments were conducted according to (i) the Plackett-Burman design (Minitab18 program) to screen several significant process factors and (ii) Central Composite design (design-Expert 11. 1. 2. 0 program) to find out the optimum process conditions for the maximum capacity of metal removal efficiency. Results: The optimum pH, metal, and bacterial concentration were 6. 76, 391 mg. L-1, and 4. 60 g. L-1 for lead removal ability of L. acidophilus ATCC4356. A quadratic model was developed to correlate the variables with removal efficiency. According to the results, this model was not statistically significant (P > 0. 05). Conclusions: The experimental removal efficiencies at the optimum condition for lead by L. acidophilus ATCC4356 (73. 9%) were consistent with the predicted values. Consequently, due to their appreciate efficiency and the lower cost of the lead removal ability, these two bacteria may be a candidate as good biosorbents. The results also confirmed that the response surface Methodology is an appropriate methodology for modeling of removal efficiency.