Background One of the problems of diabetic patients is the formation of ulcers in the lower areas, especially the legs, and the treatment of these ulcers is very important. In wounds, grampositive and Gram-negative opportunistic and biofilm-forming bacteria accumulate, and these bacteria often become resistant to antibiotics over time. For this reason, researchers are looking for new alternatives to break the barrier of antibiotic resistance. Due to their unique properties and natural origin, antimicrobial peptides can be a suitable alternative to common antibiotics. Methods In this research, several antimicrobial peptides of animal origin were investigated using docking and molecular dynamics techniques. Then, the antibacterial disk diffusion test was performed and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were measured. Results According to the Insilico results, the melittin peptide, which is present in Apis mellifera bee venom, was selected as the best candidate for measuring the antibacterial effect on the Gram-positive bacterium Staphylococcus aureus and the Gram-negative bacterium Pseudomonas aeruginosa. Then, a wound dressing was prepared using Apis mellifera bee venom, and an antibacterial disk diffusion test and the minimum inhibitory concentration (MIC) and the minimum bactericidal concentrate (MBC) measurements were performed. The results of the invitro tests showed that compared with the antibiotic ciprofloxacin, the wound dressing containing melittin could effectively inhibit the target bacteria and create a non-growth halo with an acceptable diameter. Conclusion According to the results of this research, wound dressing containing the antimicrobial peptide melittin can be a suitable option for treating diabetic foot ulcers. The inhibitory effect observed on Pseudomonas aeruginosa and Staphylococcus aureus bacteria can be generalized to all Gram-positive and Gram-negative bacteria present in diabetic foot ulcers.