In this study, we selected Gardeniae fructus (GF) as an anti-inflammatory functional material and improved the biological activity of GF through the treatment of β-glucosidase. For the simple evaluation of anti-inflammatory activity, the inhibitory activity of GF extract (GFE) on the production of NO by RAW264.7 cells in the presence of LPS was examined. β-glucosidase originating from Aspergillus niger or Aspergillus fumigatus has effectively improved the antiinflammatory activity of GFE. The enzyme treatment raised the activity of GFE by more than 10 times. The optimum conditions for the enzyme reaction were at pH 4.6, 45℃, and 20 U/mL for 24 h with agitation. In addition, in vitro production of cytokines (IL-1β, IL-6, TNF-α), COX-2, and the NF-κB activation of RAW264.7 cells decreased more in the presence of GFE treated with β-glucosidase originating from Aspergillus niger (GFAN) than in the presence of GFE. These results suggest that enzyme-treated GFE might be a potential candidate for natural anti-inflammatory food materials.