Iron (Fe) and Manganese (Mn) are essential for bacterial survival and virulence as they are involved in catalysis, infection and biofilm formation. The basal metabolism of pathogenic bacteria would be disturbed by the deficiency of metal ions in the host environment. In this study, we found that Streptococcus pneumoniae (S. pneumoniae) can still grow slowly in the medium without manganese ions. Further ICP-MS determination showed that the iron concentration in the bacterium was increased when the manganese content was decreased. The supplement of iron in the manganese deficient culture medium (MDCM) can recovery the bacterial growth. The quantitative proteome using stable-isotope dimethyl labeling was performed to investigate the adaptive growth mechanism of S. pneumoniae in Mn-deficiency condition. Compared with the bacteria cultured in the normal medium, 25 proteins were down-regulated and 54 proteins were up-expressed with more than 1.2-fold alteration (P < 0.05) in S. pneumoniae cultured with MDCM. A large number of depressed proteins are participated in cell energy metabolism, amino acid synthesis process and oxidation products reduction process. The subsequent detection indicated that both intracellar and extracellular hydrogen peroxide levels of in cells were significantly elevated, and the intracellular ATP levels of were evidently decreased in cells cultured in the absence of manganese, meaning that Mn-deficiency can cause multiple metabolic disorders. More importantly, several iron ABC transporters, such as PiuA/PiaA/PitA/SPD_1609, etc, were over-expressed to uptake more iron from the medium. These results suggest that lacking of manganese only disturbed multiple metabolic processes of S. pneumoniae, but also caused the compensatory effect of iron for manganese Mn which is beneficial to the survival of bacteria in extreme environments.