Lactobacillus contribute to maintain the human healthy and use for nutritional additives as probiotics. In this study, a cholesterol-lowering bacterium, Lactobacillus gasseri TF08–1, was isolated from the feces of a healthy adolescent, and its probiotic potentials were evaluated through genomic mining and in vitro test. The assembled draft genome comprised of 1,974,590 bp and was predicted total of 1,940 CDSs. The annotation of the genome revealed that L. gasseri TF08–1 harbored abundant categories of functional genes in metabolic and information processing. Moreover, strain TF08–1 has capacity to utilize D-Glucose, Sucrose, D-Maltose, Salicin, D-Xylose, D-Cellobiose, D-Mannose, and D-Trehalose, as the carbon source. The safety assessment showed strain TF08–1 contained few antibiotic resistance genes and virulence factors and was only resistant to 2 antibiotics detected by antimicrobial susceptibility test. A high bile salt hydrolase activity was found and a cholesterol-reducing effect was determined in vitro, which the result showed a remarkable cholesterol removal capability of L. gasseri TF08–1 with an efficiency of 84.40 %. This study demonstrated that the strain showed great capability of exopolysaccharide production, and tolerance to acid and bile salt. Therefore, these results indicate that L. gasseri TF08–1 can be considered as a safe candidate for probiotic, especially its potential in biotherapeutic for metabolic diseases. •Analysis of metabolic end products revealed TF08-1 produced acetic acid, lactic acid, and succinic acid.•TF08-1 contained few antibiotic resistance genes and virulence factors.•Helveticin J, a bacteriocin gene cluster was detected in the genome of TF08-1.•TF08-1 was found to be high tolerant to acid and bile salt concentrations.•High bile salt hydrolase activity and cholesterol-lowering activity were determined by gene prediction and in vitro test.