The soil microbial component plays a critical role in maintaining the soil functionality, performing several vital functions such as soil formation, decomposition of dead and decayed organic matter, circulation of macro and micro elements and transformation of toxic chemicals into non-toxic forms. Various Bacillus species, including B. megaterium, are known to fix atmospheric nitrogen. To ensure nitrogen fixation and diversity analysis of diazotrophs, the most widely used gene marker is nifH, which is responsible for synthesis of proteins of the nitrogenase complex. Various soil microorganisms, including B. megaterium, are endowed with the ability to solubilize phosphorus inaccessible form to accessible one and / or mineralize organic phosphorus to accessible phosphorus form. Phosphatases and phytases are two groups of enzymes that catalyze transformation of organic phosphates to inorganic ones. It was found that Bacillus megaterium exhibits acidic and alkaline phosphatase activity and produces phytases. The aim of this work was to determine by RT-PCR method the presence of genes responsible for enzyme synthesis of phytase, nitrogenase, and alkaline phosphatase in strains of previously isolated bacteria of Bacillus megaterium species. To specify the presence of genes encoding synthesis of the desired enzymes in Bacillus megaterium, an in-silico analysis of the annotated genomes of this bacterial species, presented in the NCBI information database, was carried out. Further on, the selection of primers for screening the target parts was made. According to the results of the study, 7 out of 16 isolated B. megaterium strains contained all three required DNA parts responsible for synthesis of phytase, nitrogenase, and alkaline phosphatase enzymes.