In order to enrich lactic acid bacteria resource from soy sauce residue, Lactococcus lactis from soy sauce residue were screened and identified by physiological and biochemical tests and 16S rRNA gene sequence analysis, and their tolerance to gastrointestinal environment were further studied. Four strains of lactic acid bacteria were isolated from soy sauce residue. Three of them were identified as Lactococcus lactis subsp. hordniae, HT127, HT223 and HT227, respectively. One was identified as Lactococcus lactis subsp. lactis and named HT122. The results of growth curve showed that the growth of four strains were similar, and the logarithmic growth period was 2~12 h after inoculation. The results of gastrointestinal fluid tolerance test showed that strains HT233，HT127，HT227，HT122 could tolerate the simulated gastric juice, with live bacterial rates of (94.85±0.05)%, (81.67±0.25)%, (77.99±0.58)%, (58.57±0.57)% respectively. Meanwhile, the live bacterial rates of HT122, HT127, HT223, HT227 were (77.78±0.68)%, (67.42±0.72)%, (34.25±0.08)%, (22.28±0.18)% after cultured for 8 h in the simulated intestinal juice. All the results suggested that HT127 might show the good tolerance to the simulated gastrointestinal environment and could be benefit to the expansion of food-borne lactic acid bacteria resource bank.