A microbial process for the degradation of propionitrile by Klebsiella oxytoca was studied. The microorganism, K. oxytoca, was isolated from the discharged wastewater of metal plating factory in southern Taiwan and adapted for propionitrile biodegradation. The free and immobilized cells of K. oxytoca were then examined for their capabilities on degrading propionitrile under various conditions. Alginate (AL) and cellulose triacetate (CT) techniques were applied for the preparation of immobilized cells. The efficiency and produced metabolic intermediates and end-products of propionitrile degradation were monitored in bath and continuous bioreactor experiments. Results reveal that up to 100 and 150 mM of propionitrile could be removed completely by the free and immobilized cell systems, respectively. Furthermore, both immobilized cell systems show higher removal efficiencies in wider ranges of temperature (20-40 degrees C) and pH (6-8) compared with the free cell system. Results also indicate that immobilized cell system could support a higher cell density to enhance the removal efficiency of propionitrile. Immobilized cells were reused in five consecutive degradation experiments, and up to 99% of propionitrile degradation was observed in each batch test. This suggests that the activity of immobilized cells can be maintained and reused throughout different propionitrile degradation processes. A two-step pathway was observed for the biodegradation of propionitrile. Propionamide was first produced followed by propionic acid and ammonia. Results suggest that nitrile hydratase and amidase were involved in the degradation pathways of K. oxytoca. In the continuous bioreactor, both immobilized cells were capable of removing 150 mM of propionitriles completely within 16 h, and the maximum propionitriles removal rates using AL and CT immobilized beads were 5.04 and 4.98 mM h(-1), respectively. Comparing the removal rates obtained from batch experiments with immobilized cells (AL and CT were 1.57 and 2.18 mM h(-1) at 150 mM of propionitrile, respectively), the continuous-flow bioreactor show higher potential for practical application. (C) 2010 Elsevier B.V. All rights reserved.