Antimicrobial biomaterials play important role in tissue engineering applications to protect damaged tissue from infections. The aim of this study is producing antimicrobial polycaprolactone (PCL) membranes by using a plant based antimicrobial agent. Therefore, Melissa officinalis essential oil (MEO) was investigated against 10 types of microorganisms and remarkable antimicrobial activity was demonstrated. PCL:MEO membranes were prepared by solvent casting method by mixing MEO into PCL in various ratios (PCL:0M, PCL:0.25M, PCL:0.5M, and PCL:1M w/w). Water contact angle measurements showed that hydrophilicity of the membranes increased with increasing concentrations of MEO from 103.44o to 83.36o for PCL:0M and PCL:1M, respectively. It was determined that there was an inverse relationship between the MEO concentration and the mechanical properties. Notable antioxidant activity of PCL/MEO membranes was exhibited by the inhibition percent of DPPH which was increased from 24.74% to 44.79% for PCL:0M and PCL:1M, respectively. The antimicrobial activity of MEO was also highly maintained in PCL membranes. For PCL/MEO membranes, at least 99.9% of microorganisms were inhibited. Cytocompatibility of the membranes were investigated by resazurin assay, SEM analysis and DAPI staining. PCL:0.25M and PCL:0.5M membranes supported the viability of L929 cells more than 87% when compared to PCL:0M membranes on day 6. However, the viability of L929 cells on PCL:1M membranes was about 43% indicating significant decrease on cellular activity. In conclusion, PCL:0.25M and PCL:0.5M membranes with their high antimicrobial activity, acceptable mechanical properties and cytocompatible properties, they can be considered as an alternative biomaterial for tissue engineering applications.