Effects of twelve fermentation media, temperature, pH and culturing time on nicotine degradationby bacteria strain 4 # and 9#were studied. Results showed that 11# medium was suitable for the growth of strain4#, and 5#, 7# and 9# medium were suitable for nicotine degradation. Suitable medium for growth of strain 9# was8# medium, and the ability to degrade nicotine was higher in medium 7#, 9#, 11# and 12#. Suitable culturing conditions for strain 4 # and 9# were at 30℃, pH 6.5,48h and 35℃, pH 6.5, 56h respectively.