Abstract: In order to evaluate the safety and probiotic properties of Lactobacillus plantarum HCS03-001(CGMCC No.16258), the safety of Lactobacillus plantarum HCS03-001 was evaluated by hemolysis test, drug resistance test and acute transoral toxicity test. The probiotic properties of the strain were evaluated by acid and bile salt tolerance test, bacteriostatic functional properties test and laxative efficacy test. The results showed that Lactobacillus plantarum HCS03-001 did not form transparent circle on the Colombian blood finger culture medium, that was, there was no hemolysis. Lactobacillus plantarum HCS03-001 was sensitive to penicillin (PEN), ampicillin (AM), imipenem (IP), meropenem (MP), linezolid (LZ), and datoramycin (DPC), and neutral to erythromycin (EM) and resistant to vancomycin (VA). The acute oral toxicology of mice was non-toxic, and the strain was confirmed to be safe to eat.The survival rate of the strain was 94.29% and 92.33% at pH3.0 and pH2.0 for 3 h, respectively. The survival rate could still reach 88.82% at 1.5 g/L bile salt for 3 h. It had inhibitory effect on intestinal and oral pathogens (Porphyromonas gingivalis and Streptococcus mutans). It had a good effect on promoting defecation in constipation model of loperamide hydrochloride. Lactobacillus plantarum HCS03-001 has good safety and probiotics.