The relationship between bioluminescence of immobilized Photobacterium phosphoreum and toxic substances was investigated to monitor toxic substances in aqueous solution. The sodium alginate was used as an immobilization matrix. A bioluminescence intensity was maximum when OD660 for cell concentration were between 1.0 and 1.2 and the biolumescence was stable at the pH range of between 6.0 and 8.0. The optimum concentration of alginate for immobilization was found to be 5.0%(w/v) in which dilution was carried out with 2.5%(w/v) NaCl solution that is an optimum environmental condition for the growth of P. phosphoreum. The bioluminescence intensity responded against the toxic substances was proportional to the concentration and a regression curve were established with linearity by using specific bioluminescence reduction rate and Gamma values. It was also found that the response was very rapid and sensitive. The response with such rapidity and sensitivity is a very important factor for the real time monitoring. The immobilized cells showed higher sensitive response to the toxic substances than free cells.