Media is an important substrate for the growth of different types of microorganisms. Themost common growthmedia formicroorganisms are nutrient broth and agar plates. The aimof the investigationwas to reutilize the medium by supplementing the lost constituents, thereby reducing the investment on media. The protein and carbohydrate content decreased when themediumwas reutilized for culturing proteolytic bacteria and fungi. The growth of the microorganisms was good when the media was resupplemented with lost nutrients such as protein and carbohydrate when compared with the media which was not resupplemented. The effect of media supplemented with deficient nutrients was studied on proteolytic bacteria and fungi, the growth of the microorganisms decreased after supplementation of themedia when compared to growth on freshmedium.