Ezine cheese is a nonstarter and long-ripened cheese produced in the Mount of Ida region of Çanakkale, Turkey, with a protected designation of origin (PDO) status. The nonstarter fermented foods serve as sources for the isolation of novel strains. The present study aimed to report the novelty of the bacteriocinogenicEnterococcus lactisPMD74 strain and thein vitroassessment of its potential as a probiotic candidate. Additionally, the present study aimed to describe the technological and safety-related properties of the aforementioned strain. The strain exhibited high viability at pH 3.0, in the presence of pepsin, pancreatin, and bile salts (0.3% and 0.5%), and considerable survival passage through the stimulated digestion tests. The strain PMD74 exhibited substantial autoaggregative (41%) and coaggregative properties, which increased as a function of time. The highest coaggregation percentage was obtained withSalmonella entericaserotype Typhimurium SL1344 (23%), followed byStaphylococcus aureusATCC 6538 (10.3%) andEscherichia coliATCC 26922 (7.4%), respectively. The strain PMD74 was able to inhibit the growth of a number of Gram-positive bacteria, includingListeria monocytogenes,Lactobacillus sake,Staphylococcus aureus, andE. faecalis. The antimicrobial activity of the proteinaceous compound was calculated as 6400 AU·mL−1by the critical dilution method againstE. faecalisATTC 29212.γ-Hemolytic PMD74 was observed to be sensitive to vancomycin, ampicillin, penicillin, gentamicin, tetracycline, chloramphenicol, and tylosin. Among the four genes tested,E. lactisPMD74 was observed to be positive for three virulence determinants,ace,sprE, andgelE, and negative foresp. The amino acid decarboxylase activities were detected negative for histidine, tyrosine, and ornithine.E. lactisPMD74 was classified as a low acidifier, which suggested its possible role as an adjunct culture.E. lactisPMD74 exhibited considerable survival ability (8.86 log CFU·mL−1) in the acidic condition of fermented milk for a four-week-long storage period.