The aims of this work were to characterize and determine bioactivities of crude exopolysaccharide (EPS) extract from Bacillus tequilensis PS21 isolated from milk kefir from Kampaeng Petch, Thailand. B. tequilensis PS21 produced 112.1 mg dried EPS/l from initial 80 g/l lactose in modified TSB media at 52 h, with EPS product yield of 8.9 mg EPS/g lactose and specific product yield of 0.3 mg EPS/mg biomass. The FTIR result confirmed EPS to be a protein-bound polysaccharide and SEM analysis showed the morphology to be a grainy appearance with an uneven surface, covered with pores. HPLC analysis determined EPS as a heteropolysaccharide consisting of five sugar units with the following molar ratios; xylose (17.65), glucose (2.54), ribose (1.83), rhamnose (1.23), and galactose (1). Chemical components of this EPS were predominantly carbohydrate at 697.8 mg/g EPS (65%), protein 361.4 mg/g EPS (34%), and nucleic acid 12.5 mg/g EPS (1%). The EPS demonstrated antioxidant activities at 57.5% DPPH scavenging activity, 37.2 μM Fe(II)/mg EPS and 34.9 μM TEAC μM/mg EPS using DPPH, FRAP and ABTS assays, respectively. EPS also exhibited anti-tyrosinase activity at 34.9% inhibition. This work represents the first finding of EPS produced by Bacillus sp. from Thai milk kefir which shows potential applications in the production of antioxidant functional foods and whitening cosmetics. However, optimization of EPS production for industrial exploitation requires further study to ascertain the economic potential.