A new technology for 6-hydroxy-3-succinoyl-pyridine (HSP) production from (S)-nicotine in tobacco waste by whole cells of a Pseudomonas sp. has been developed. When deionized water was used in the transformation reaction as a medium and the initial pH value of reaction mixture was adjusted to 7.0, 1.45 g/L HSP was produced from 3 g/L of nicotine in 5 h with 3.4 g/L of cells in a 5-L flask at 30 degrees C. HSP could be easily purified from the reaction without perplexing separation steps. A quantity of 1.3 g of HSP was recovered without impurity, and the overall yield of HSP was 43.8% (w/w), based on an initial concentration of 3.0 g/L of nicotine in reaction. This biotransformation made it possible to convert nicotine in tobacco wastes with high nicotine content into valuable compounds.