Curd is a source of probiotic Lactobacilli. In the present study, ten homemade curd samples were collected from different regions of Vellore district, Tamil Nadu, India. Among the samples, four different strains of Lactobacillus sp. were isolated and identified based on their colony morphology and biochemical characteristics. It was observed that isolated Lactobacillus spp. were resistant to inhibitory substances like NaCl (1-9%), bile-salt (0.05-0.3%), and showed good growth in the acidic condition, while maximum growth was observed at pH around 6. The isolates were examined for their antibacterial activity against four different test pathogens, and found that growth of all pathogens are inhibited to some extent but maximum zone of inhibition was observed against E.coli (33mm) and no zone of inhibition against Pseudomonas aeruginosa after 24 hour incubation. The isolated Lactobacillus sp. showed good survival abilities in acidic (pH 4) and alkaline (pH 8) conditions. Isolated Lactobacilli were able to produce organic acid in skim milk which was determined by titrimetric method. In the antibiotic Sensitivity test, LB06, LBS3, LB02 and LBS1 were found to be sensitive against Ciprofloxacin andErythromycin. LB06 was resistant against Ampicillin and Bacitracin while LB02 was resistant againstAmpicillin. In the DPPH scavenging assay, LB06 and LBS3 showed highest inhibition of 56.84% and 55.86% in 1000 μl/ml compared to the positive control (BHT) 69.29%. The IC50 value was found to be 750μl/ml.