Abstract: The aim of this study was optimization of culture medium in direction of increasing the production rate of desferrioxamine B. Streptomycetes are the most widely studied and well known genus of the actinomycete family. Streptomycetes usually inhabit soil and are important decomposers. The genus Streptomyces are Gram-positive and GC rich bacteria that are important for production of many antibiotics and secondary metabolites. These metabolites are important in industrial and medical fields. Deferoxamines (also known as desferrioxamine B, desferoxamine B, DFO-B, DFOA, DFB or desferal) are low-molecular-weight, iron-chelating compounds (siderophores) produced and secreted by many actinomycetes, including species of Streptomyces, Nocardia and Micromonospora. Streptomyces pilosus synthesizes the siderofore desferrioxamine B. Desferrioxamine B is used clinically to treat disorders related to iron overload and pathological iron deposition in human. Our results revealed that the use of soybean as a base medium plus additives such as Na2HPO4.12H2O, NaH2PO4, MgSO4.7H2O, ZnSO4.7H2O, FeSO4.7H2O, CaCl2.2H2O, NaCl, MnSO4, NH4Cl, KH2PO4, K2HPO4, some of the amino acids and vitamins increased the production of desferrioxamine B about 8 times in comparison with the control.