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Lactic acid bacterium isolated from fermented durian (tempoyak) was investigated for its potentials as a probiotic strain. Bacterial tolerance toward gastrointestinal environment, adhesion, and cytotoxic activity in human colon adenocarcinoma cell line HT-29 was evaluated. 16S rRNA sequencing identified the lactic acid bacterium as Lactobacillus plantarum . The bacterium demonstrated good tolerance toward gastrointestinal pH 2.0 and 0.3% bile salts. It showed strong adhesive capacity in human intestinal cell line, HT-29, with an adhesion index of 159 ± 10. Cytotoxicity of L. plantarum was investigated using both live bacterial cells (BC) and cell-free supernatant (CFS). Findings showed that both BC and CFS of L. plantarum reduced proliferation of HT-29 colon adenocarcinoma cells using MTT assay. The results imply potential probiotic properties of L. plantarum isolated from tempoyak.