Strain NK34 was characterized for probiotic use. Strain NK34 was named Lactococcus lactis NK34 based on API 50 CHL kit results and 16S rDNA sequencing. L. lactis NK34 was highly resistant to artificial gastric juice (pH 2.5) and artificial bile acid. Based on results from the API ZYM kit, 4 enzymes were produced. L. lactis NK34 was resistant to all antibiotics tested except for $10;{mu}g/mL$ roxithromycin and $10;{mu}g/mL$ erythromycin. The cholesterol-lowering effect of L. lactis NK34 was about 46.9%. Concentrations of interleukin $(IL)-1{alpha}$ in the $20{ imes}$ concentrated supernatant of L. lactis NK34 was about 361 pg/mL. L. lactis NK34 was also found to inhibit the growth of colon cancer cells due to MNNG-induced DNA damage. These results demonstrate the potential of L. lactis NK34 as a health-promoting probiotic.