Research on the association of serum folate levels with metabolic syndrome (MetS) in premenopausal women is lacking. This study was aimed to investigate this association in 1730 premenopausal women using the 2016 to 2018 Korean National Health and Nutrition Examination Survey data. Participants’ mean age and BMI were 35.9 years and 22.7 kg/m2, respectively. Participants were divided into three groups according to serum folate tertiles. Odds ratios (OR) and 95% confidence intervals (CIs) for abdominal obesity, elevated blood pressure (BP), high fasting plasma glucose (FPG), high triglycerides (TG), low high-density lipoprotein cholesterol (HDL-C), and MetS were calculated in multiple logistic regression models adjusted for possible confounders, by serum folate level tertiles. Prevalence of MetS (14.9, 11.0, and 8.6%); abdominal obesity (17.8, 16.0, and 11.4%); high TG (17.5, 14.0, and 11.1%); and low HDL-C (50.3, 44.6, and 42.5%) decreased with increasing folate level tertile. Prevalence of elevated BP (14.3, 12.0, and 11.7%) and high FPG (11.9, 15.8, and 13.0%) showed no significant differences according to serum folate level tertiles. The multivariate-adjusted ORs (95% CIs) for MetS, abdominal obesity, elevated BP, high TG, and low HDL-C in the highest folate level tertile were 2.17 (1.46–3.22), 1.80 (1.25–2.60), 1.77 (1.16–2.70), 1.90 (1.35–2.67), and 1.49 (1.14–1.94), respectively. The ORs for high FPG did not show significant differences according to serum folate level tertiles. In conclusion, serum folate levels were inversely associated with an increased risk of MetS in Korean premenopausal women. These results suggest that MetS can be prevented and managed by improving the serum folate levels in premenopausal women.