ABSTRACTDecaffeination of food and beverage products is in high demand. In this study, a caffeine-degrading bacterium Burkholderia spp. was isolated from coffee plantation area of Chiang Mai province of Thailand. The bacterial isolates were first identified by morphological, physiological, and biochemical tests followed by 16S rDNA analysis. The bacterial isolate of Burkholderia spp. showed 45.5% of caffeine degradation in caffeine containing media (2.5 g/L) after 110 h of incubation period. Burkholderia spp. showed only 2.6% caffeine degradation when exposed to high concentrations of caffeine containing medium (20 g/L). The growth rate of Burkholderia spp. declined with the increase in the caffeine concentration, which indicated the inhibiting effect of caffeine at very high concentrations. The maximum growth rate of 0.053 h−1 was observed at 2.5 g/L of caffeine. Overall due to high caffeine tolerance and biodegradation of caffeine, Burkholderia spp. can be effectively used to degrade caffeine from agro-industrial wastes targeted for value added food applications and environmental remediation.