Pseudomonas arsenicoxydans has been recently described as a new arsenite oxidizing bacterial species. Arsenite detoxification activity by this species was determined by HPLC/HG/AAS. P. arsenicoxydans showed a high rate of As(III) conversion, particularly when immobilized (it oxidizes 100 % of 500 μg arsenite present in the medium after 48 of incubation). Arsenite oxidizing activity, mediated by a constitutive periplasmic enzyme, was determined following the transfer of reducing equivalents from arsenite to 2,4-dichlorophenolindophenol (DCIP) showing that approximately 75 % (0.173 µmol DCIP min(-1) mg(-1)) of the total activity (0.231 µmol DCIP min(-1) mg(-1)) was detected in the periplasmic fraction. Using PCR with primers specific for arsenite oxidase gene showed the presence of a gene encoding for arsenite oxidase in P. arsenicoxydans. Results show the potential biotechnological application of P. arsenicoxydans as a candidate for detoxification of As(III).