Since the condition of immobilization must be optimized, it is very important to know whether and on how conditions bacterial cells retain their metabolic activity during immobilization process. A bioluminescence intensity had the maximum value when cell concentrations were between 1.0 and 1.2 measured at O.D660. The strontium alginate was used as an immobilization matrix and two independent factors for immobilization of Photobacterium phosphoreum with strontium alginate were optimized with the response surface methodology(RSM) considering degree of bioluminescence. As a result, the optimum concentration for immobilization was found to be 2.4%(w/w) for sodium alginate and 0.31M for strontium chloride, respectively. A dilution was carried out with 2.5%(w/v) NaCl solution that is an optimum environmental condition for growth of P. phosphoreum. Under the such condition of immobilization, hardness could be predicted as 4.66$ imes$104N/$m^2$ and it took different time according to the volume of matrix to be immobilized completely.