The aims of this research were to apply the sourdough LAB containing specific starter cultures for Barbari bread production and evaluation it`s microbiological shelf life and sensory properties. The effects of fermentation times (8, 16 and 24 h), fermentation temperatures (28, 32 and 36 °C) and type of starter culture (Lactobacillus sanfransicencis (ATCC 14917), Lactobacillus plantarum (ATCC 43332) and a mixture of both LAB) were analyzed in a completely randomized design with factorial experiment and 4 replications was conducted. Microbiological shelf lives of samples were determined by plating serial dilutions on to plate count agar, at days 1 to 7. Breads sensory properties were determined in 1, 24, 48 and 72 h intervals, after baking according to AOAC standard method. The results showed that, sourdough had significant effect (p≤0.05) on microbiological shelf life and sensory properties of Barbari bread in comparison with control sample. Moreover the produced sample with Lactobacillus plantarum (24 h fermentation time and 32 °C fermentation temperature) had the most microbiological shelf life and the sample produced with a mixture of both LAB (16 h fermentation time and 28 °C fermentation temperature) had the best sensory evaluation, 72 h after baking.