From a rheological and physical perspective, protein-starch-fat interactions could be examined through imitation cheese. Pregelatinized starches (native and modified) were used to partially replace protein and fat in the production of imitation cheese and were compared with control imitation cheese (0% starch). The resultant imitation cheese replaced with pregelatinized starches were softer, more cohesive, and improved melting properties compared to control. The dominancy of the elastic component (G′ > G″) was confirmed by dynamic rheological analysis. All samples demonstrated pseudoplastic behavior (i.e., flow behavior index < 1). Inclusion of chemically modified pregelatinized starches (lactic and citric acid) in imitation cheese showed storage moduli (G′) near to control. These distinctive properties in the present study could be helpful to control the rheological and physical characteristics of imitation cheese by proper concentration of casein, fat, and modified starches. Practical applications Low calorie and cheap imitation cheese can be prepared with improved functional and rheological properties by partially replacing casein and fat with pregelatinized starch citrates and lactates. These pregels allowed easy simultaneous replacement of casein by 15% and fat by 10% in imitation mozzarella cheese. How to cite this article: Butt NA, Ali TM, Hasnain A. Development of rice starch-based casein and fat mimetics and its application in imitation mozzarella cheese.