In this work, the parameters affecting the recovery of copper from the low-grade sulfide minerals of Sarcheshmeh Copper Mine were studied. A low-grade sulfide ore was used with a copper grade of 0.25%, which was about 28% of the mineral oxide, and the sulfide minerals made up the rest. Much more sulfide minerals were found to be pyrite and most of the gangue minerals were quartz, anorthite, biotite, and muscovite. In order to investigate, simultaneously, the solids (10 to 20%) and acidity (1.5 to 2.5) and shaking (110 to 150 rpm), the separation of bacteria from Sarcheshmeh Copper Mine was carried out. After adjustment of the sample, bio-leaching tests were performed in accordance with the pattern defined by the software DX7 in shaking flasks, and the Cu recovery was modeled and optimized using the response surface methodology. The influential parameters were comprehensively studied. The central composite design methodology was used as the design matrix to predict the optimal level of these parameters. Then the model equation was optimized. The results obtained showed that increasing solids (from 10 to 20%) was bad for bacteria. The highest copper recovery was equivalent to 69.91%, obtained after 21 days at 35 degrees using the Acidi Thiobacillus Ferrooxidans bacteria and a K9 medium with a pulp density of 10% and pH 1.5.