Nanofibrous zein/PEG based membranes incorporated with natural antimicrobial compounds were fabricated by electrospinning method. Structural and thermal analysis of prepared nanofibers revealed that the applied processing technique did not significantly affect the structure of pristine zein polymer. Morphological characterization showed a higher degree of polydispersity in the fibers modified with eugenol, thymol, nisin, or their combinations, and an average fiber diameter in the range from 300 to 390nm. Nanofibrous samples with eugenol and thymol prevented the growth of Escherichia coli and Staphylococcus aureus, while the nisin or its mixtures with phenols proved a high antibacterial effect against Gram-positive Listeria ivanovii. Zein/PEG membranes with bioactive molecules significantly eliminated biofilm formation, with the most pronounced effect of zein/PEG/Eug/Thy combination. Biodegradability testing of bioactive membranes revealed no significant slowdown of degradation process in comparison to control sample. Zein/PEG hydrophilic nanofibers enriched with phenol/nisin combinations demonstrated a high potential for development of sustainable packaging to improve the shelf-life and quality of foods.