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A novle nicotine-degrading bacterrium, strain, was isolated and identified as Pseudomonas putida strain based on its 16S rDNA sequence and morphological and biochemical features. The isolate could utilize as the sole source of carbon and nitrogen. Nicotine (3.0g/L) was degraded by Pseudomonas putida strain A3 within 48 h at 28 and pH 7.0. Therefore, Pseudomonas putida A3 has the potential to degrade nicotine during tobacco leave processing.