Secondary metabolites are the special chemicals that organisms make for their own purposes. They are not needed for their basic functions, but they have many uses in their interactions with the environment. They can protect themselves, communicate with others, compete with rivals, and cooperate with partners. Mycorrhizal Helper Bacteria (MHBs) are bacteria that enhance the plant-fungi partnership by supporting their growth, nutrition, and defense, and by modifying their production and use of secondary metabolites, which help them interact with the environment. In this study, morphological characterization and isolation were carried out following the serial dilution method, and checked the antimicrobial activity of isolated strains in biological screening. The best strains were selected for secondary metabolite production through shaking fermentation culture techniques; FTIR (Fourier transform infrared) spectroscopy and grams’ staining were also carried out. Strain PW 2-3-1 showed the highest antimicrobial activity, whereas strain AP 10-2-4 showed the least against four bacterial strains, viz; Bacillus meurellus, Bacillus subtilis, Acinetobacter rhizosphaerae, and Escherichia coli. FTIR spectrum analysis showed the presence of C–H and C–O stretches with wavenumbers ranging from 500-3500 of antibiotic nature. The application of Mycorrhization Helper Bacteria can be an encouraging method to achieve successful reforestation. It has been additionally recommended that Mycorrhization Helper Bacteria could detoxify the impacts of parasitic metabolites.