Rapamycin (Rap), as a macrocyclic polyketide with immune suppressive, antifungal, and anti-tumor activity produced by Streptomyces hygroscopicus, is receiving considerable attention for its significant contribution in medical field. However, the production capacity of the wild strain is very low. Due to the high price of rapamycin drug (1MG 30/TAB price= 900PT) and the limited number of producing organisms, the present research was conducted to search for novel strains from local environments capable of producing rapamycin. Streptomyces sp. RHS5 selected from thirty-three actinobacterial isolates was the most potent in producing the active metabolite and was compared with rapamycin produced by Streptomyces hygroscopicus using bioautography. The chemical structure of the purified compound determined by spectroscopic analyses was found to be analogue to rapamycin. So far, considerable efforts have been made to improve rapamycin production, such as fermentation technology optimization. In this research, the production of rapamycin analogue was economically achieved by applying the solid-state fermentation (SSF) using wheat bran as a substrate and optimization of culture condition was achieved by the statistical design Plackett- Burman. The productivity of rapamycin analogue after optimization increased by 1.18 fold compared to basal medium.