The objective of this study was to investigate probiotic, antimicrobial, technological and safety properties of lactobacillus strains isolated from local Iranian cheese made from raw milk. Six different samples were prepared, after serial dilution, culture was performed on MRS culture medium. The gram‐positive and catalase‐negative lactobacillus strains were subjected to grouping and identifying using biochemical tests, carbohydrates fermentation profiles, and 16S rDNA analysis. The results of sequence analysis showed the Lactobacillus spp. belonged to Lactobacillus brevis, Lactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus casei. After 3 hr incubation at pH=2, 3–6 log units of strains decreased which Lactobacillus acidophilus (B14) and Lactobacillus brevis (B2) showed highest resistance to low pH as well as simulated GIT juices. The highest and lowest hydrophobicity degree was belonged to L. acidophilus (B14) (65.9%) and L. casei (B22) (25.6%), respectively. Also, the highest auto‐aggregation and coaggregation were observed in L. acidophilus (B14) (51.3%) and L. plantarum (B20) (43.6%). The adhered percentage of strains varied from 2.5% to 14.6%. L. plantarum (B20) showed highest proteolytic activity followed by L. acidophilus (B14). Also, the highest autolytic activity belonged to L. acidophilus (B14). All of the strains showed low acidifying potential, except for L. acidophilus (B17) which decreased 2.05 unit of pH after 24 hr. The isolates did not show lipolytic activity as well as biogenic amines production (except L. brevis B3). All of the strains were sensitive to chloramphenicol and erythromycin except L. acidophilus (B15) and L. casei (B22). All strains showed no hemolysis activity which make them safe for consumption. Based on the obtained results, L. acidophilus (B14) presented the best probiotic and technological characteristics and is proposed for using as coculture in the dairy industrial.
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