Lactic acid bacteria are very significant to human health due to the production of some antimicrobial substances and ability to inhibit pathogenic bacteria. Furthermore, the bacteria are also used as starter culture in the production of various foods. The present study was focused on isolation and characterization of Lactobacillus acidophilus from yogurt and to demonstrate some of probiotic properties of these isolates. All isolates were phenotypically characterized including studying, biochemical, effect of sodium chloride and pH during growth, carbohydrates test and characterizing the antimicrobial activity of Lactobacillus acidophilus against pathogens. The present study demonstrates that Lactobacillus acidophilus produced a bacteriocin- like inhibitory substance with a broad spectrum of antimicrobial activity directed against pathogenic indicator organism suggesting its protective value against enteric pathogens.