This study aimed to develop a Cheddar cheese using biological preservatives as substitutes for chemical preservatives. Nisin, lysozyme and lactic acid bacteria (Lactobacillus rhamnosus) were used as biological preservatives while NaNO3 was used as a chemical preservative. Physico-chemical properties of cheese such as pH, titratable acidity, texture and moisture were measured during four weeks of ripening storage at 16 °C. Sensory properties of cheese were evaluated with the help of untrained sensory panel using a five point hedonic scale and total lactic acid bacteria, yeast and mold and coliform counts were also determined. In nisin incorporated samples, mean sensory scores for flavor, texture, odor and overall acceptability were higher than other cheese samples (p