PurposeThis research intended the utilization of Javanese turmeric (0.4% w/v) as a kombucha substrate and analysis of its hepatoprotective activity, in comparison against nonfermented Javanese turmeric beverage (JTB) and black tea kombucha.Design/methodology/approachForty-two healthy male Balb/c mice (two- to three-week-old, 20–30 g) were divided into six groups with seven replicates each. The treatments were normal diet, normal diet + Javanese turmeric kombucha (JTK), normal diet + diethylnitrosamine (DEN), DEN + JTB, DEN + JTK, DEN + black tea kombucha. Kombuchas and JTB were given at 0.3 mL/20 g BW/d. DEN was induced intraperitoneally at a dose of 100 mg/kg. Observed biomarkers were blood serum glutamate pyruvate transaminase (SGPT) and serum glutamate oxaloacetate transaminase (SGOT) activity, serum malonaldehyde (MDA), as well as liver histology. Data were analyzed using analysis of variance.FindingsAmong DEN-induced groups, JTK significantly (p < 0.05) diminished the level of blood SGPT, SGOT and serum MDA. JTK also had lower blood SGPT (8.604 ± 2.195 U/L) and serum MDA levels (2.884 ± 0.083 nM/mL) compared to the normal group (8.604 ± 2.195 U/L and 5.050 ± 0.998 nM/mL, respectively). JTK also produced the least damaged liver-cell numbers.Originality/valueJTK demonstrated better hepatoprotective activity compared to JTB.