The effects of Cd, Ni, Cu, or Zn on the whole cell-derived fatty acid profiles of four bacterial strains isolated from heavy metal-polluted soils located in Upper Silesia was determined. Based on the fatty acid methyl ester (FAME) profiles, the strains were identified and named as Enterobacter intermedius AM15, Enterobacter intermedius MH8b, Pseudomonas putida MH1d, and Klebsiella pneumoniae AM12. The obtained results showed changes that were dependent both on tested strains and metal used. The most significant changes were observed for strains cultured in the Ni presence. In the FAME profiles of MH8b, AM15, and AM12 strains, a significant increase of cyclopropane fatty acids was observed. Moreover, exposure for Ni resulted in the appearance of a new fatty acid in the FAME profiles of AM15 and MH8b strains. In turn, Cd and Zn caused a decrease of the content of cyclopropane fatty acids as compared to control. For AM15 and AM12 strains cultured on media with heavy metals, the ratio of saturated to unsaturated fatty acids were higher than that in control. The same phenomenon was also observed for MH8b strain exposed only to the highest concentration of Ni and Cd