Abstract: Utilizing nicotine as the sole source of carbon and nitrogen,152 bacterial strains were isolated from tobacco growing soils in Zibo,Shandong Province and tobacco waste from Wuhan Cigarette Factory,respectively,and a high concentration nicotine degradation bacterium strain ZB-16A was screened out from them.Studies of normal morphology,physiological and biochemical characterizing,16S rDNA gene sequence and phylogenesis tree analyses indicated that strain ZB-16A belongs in Pseudomonas putida.ZB-16A tolerates 6 000 mg/L nicotine concentration and completely degrades nicotine in 48 hours at 30℃ when initial pH of culture medium and rotation speed of shaker are 7.0 and 150 rpm,respectively;while the growth of ZB-16A is inhibited at an initial nicotine concentration of 7000 mg/L.