The object of this study is to evaluate the feasibility of microbial degradation of nicotine in aqueous extract of waste tobacco (AEWT) which was utilized for preparation of reconstituted tobacco. One novel nicotine-degrading strain was isolated from waste tobacco. Based on standard morphological, physiological assays and nucleotide sequence analyses of enzymatically amplified 16s ribosomal deoxyribonucleic acid, it was identified as Pseudomonas sp. ZUTSKD with a Genebank Database access number of EF538425. Due to the higher concentration of nicotine, NaCl, and reducing sugar in AEWT, the strain had to go through a course of adaptation. The tolerance of Pseudomonas sp. ZUTSKD to NaCl and nicotine was improved from 2.0 % to 3.5% and from 4.5 g/L to 5 g/L respectively. Its tolerance to glucose was up to a concentration of 40 g/L. The domesticated strain showed the nicotine degradation ability under higher NaCl and nicotine concentrations, even in the AEWT. In addition, it was found that the addition of glucose improved the cell growth but inhibited nicotine degradation as glucose concentration increased. It could be concluded that Pseudomonas sp. ZUTSKD could degrade the nicotine in AEWT. However, its tolerance to reducing sugars needs to be further improved.