Carotenoids are valuable molecules as they used in manufacturing of food, pharmaceutical, poultry and cosmetics of which have antioxidant properties and act as vitamin A precursors. Fusarium camptoceras ASU5 (KU215707) was selected as a promising candidate for β- carotene production. This fungus is characterized by abundant macroconidia in sporodochia with pointed apical cell and conical to point basal cell. Microconidia and mesoconidia are produced singly from monophalidic and polyphialidic conidiogenous cells. The physic-chemical characteristics of the β- carotene basal medium were screened by applying one factor. The initial screening revealed two components: molasses and (NH4)2PO4 influencing yields of β-carotene. The optimal factors were then combined for further studies through the central composite design (CCD) of response surface methodology (RSM) to optimize the significant values and to study the mutual interactions between these factors. The analysis revealed that optimal medium concentrations for maximum production of β- carotene (508.3 mg/L) were (g/L): Molasses 50; (NH4)2PO4 1.2; KH2PO4 2.5; L- asparagine 0.1 and MgSO4.7H2O 0.1. Analysis of variance results showed that the model was significant and an R2 value of 98.2% demonstrated that the experimental results were fitted well with predicted values.