In this study, a total of 22 Lactobacillus species were isolated from anchovy (Engraulis encrasichalus) samples, purchased at fish markets in Ankara-Turkey, in order to select the most suitable strains according to their metabolic and antimicrobial activities including probiotic properties. The results of identification were as follows: Lactobacillus lactis (54.5%), Lactobacillus helveticus (22.7%), Lactobacillus bulgaricus (13.6%), and Lactobacillus del-brueckii (9.1%). The different amounts of lactic acid and hydrogen peroxide produced by the Lactobacillus spp. were determined. Lactic acid levels were between 2.3-5.1 mg/mL, and all of the strains produced hydrogen peroxide. Antimicrobial effects of the Lactobacillus spp. strains on Escherichia coli K12, Staphylococcus aureus 4-43, Pseudomonas fluorescens RSKK 240 and Enterobacter aerogenes RSKK 585 were also determined by an agar diffusion method. All of the strains were able to inhibit the tested bacteria.