Few N-1-(β-D-ribofuranosyl) heterocyclic mimetics have been designed using in silico techniques, synthesized by Vorbrüggen and Bennua method and subjected to evaluate in vitro antimicrobial activity. The global reactivity descriptors were evaluated using FMO energies data to analyse the chemical properties and biological activity of compounds. Several mode of interactions, binding affinity and 50% effective concentration of compounds were computed by docking simulations against peptide deformylase (PDF) protein, using Discovery Studio v20.1.0.19295. MD simulations were also executed on the best screened compound for a better understanding of binding affinity and stability of compound within the inhibitory binding pocket of PDF by employing GROMACS 2019.6 package. Furthermore, antibacterial screening in combination approaches was carried out for all compounds using standard antibiotics chloramphenicol and kanamycin through broth micro-dilution method. A promising synergistic effect with two-fold to sixteen-fold better inhibitory potency was exhibited by compounds against different wild type human pathogenic bacteria, viz. B. cerus, B. subtilis, S. aureus, E. coli and P. aeruginosa, and one mutant strain of E. coli. Moreover, antifungal screening was also performed for all compounds against four pathogenic fungal species, viz. A. flavus, A. niger, F. oxysporum and C. albicans by two different methods and compound 3 was obtained as the best screened one. The study clearly demonstrated that in silico and in vitro results found in good coherence. Thus, the lead compound could be potentially developed as an antimicrobial agent.