A new technology for 6-hydroxy-3-succinoyl-pyridine(HSP) production from (S)-nicotine in tobacco waste bywhole cells of a Pseudomonas sp. has been developed.Whendeionized water was used in the transformation reactionas a medium and the initial pH value of reaction mixturewas adjusted to 7.0, 1.45 g/L HSP was produced from 3 g/Lof nicotine in 5 h with 3.4 g/L of cells in a 5-L flask at 30°C. HSP could be easily purified from the reaction withoutperplexing separation steps. A quantity of 1.3 g of HSPwas recovered without impurity, and the overall yield of HSPwas 43.8% (w/w), based on an initial concentration of3.0 g/L of nicotine in reaction. This biotransformation madeit possible to convert nicotine in tobacco wastes withhigh nicotine content into valuable compounds.