Ensete ventricosum (Welw.) cheeseman which belongs to the family of Musaceae is one of the main sources of starch in Ethiopia. This study aimed at evaluating epichlorohydrin cross-linked enset starch as a drug release sustaining excipient in microsphere formulations of theophylline. Extracted Enset starch was cross-linked using epichlorohydrin as a cross-linking agent. The effect of cross- linker concentration, cross-linking duration and cross- linking temperature on the degree of cross- linking and release rate of microspheres prepared by emulsion solvent evaporation method was investigated using two-level full factorial design. Accordingly, the concentration of epichlorohydrin and duration of cross-linking were the most significant factors affecting both the degree of cross-linking and drug release rate. Thus, the effects of these two factors were further studied and optimized using the central composite design. As per the numerical method of central composite design, the optimal points were obtained at epichlorohydrin concentration of 13.70% and cross-linking time of 3.82h. Under these optimal conditions, the model predicts the degree of cross-linking of 74.70% and drug release