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Objective: To investigate the probiotic characteristics, anti-Candida activity, and antibiofilm attributes of Hentak derived Lactobacillus pentosus strain LAP1., Design: The probiotic properties of strain LAP1 were depicted by adapting standard protocols. The anti-Candida and antibiofilm properties of isolate were determined using agar well diffusion assay and ELISA reader test, respectively. The time-kill assay was performed using viable colony count assay. Further, the co-aggregation property of strain LAP1 was determined based on standard methodology., Results: Strain LAP1 exhibited not only tolerance to acidic pH but also showed resistivity (P ≤ 0.05) to simulated gastric juice exposure. Similarly, the strain was able to tolerate bile salt, showed hyperproteolytic activity, and also depicted susceptibility to most of the antibiotics tested. Auto-aggregation phenomenon (37.5-60%), hydrophobicity nature (42.85%), and survival potentiality of strain LAP1 under freeze-dried condition (9.0 ± 0.01 log CFU/ml) made the isolate a promising probiotic candidate. Cell-free neutralized supernatant (CFNS) of strain LAP1 exhibited potent antifungal activities against C. albicans, C. tropicalis, and C. krusei with arbitrary unit of 150 ± 4.34, 200 ± 5.21, and 130 ± 5.13 AU/ml, respectively and depicted remarkable reduction in the biofilm formation of respective Candida sp. in a concentration dependent manner. Moreover, time-kill assay data provided the growth inhibition of all Candida sp. in a time dependent manner. Additionally, strain LAP1 revealed significant co-aggregate percentage with C. albicans, C. tropicalis, and C. krusei., Conclusions: L. pentosus strain LAP1 exhibited a good probiotic characteristics, potent anti-Candida activity, and significant antibiofilm property that could be undoubtedly recommended for its vast applications not only in food industries but also as biotherapeutic agent against Candida infections in pharmaceutical industries., (Copyright © 2018 Elsevier Ltd. All rights reserved.)