Banda Aceh city and Aceh Besar Regency are two of the leading tourism areas located in the province of Aceh. For travelling, there are some important things to be considered, such as determining schedule and distance of tourism. Every tourist certainly chooses the shortest route to reach the destination since it can save time, energy, and money. The purpose of this reserach is to develop a method that can be used in calculating the shortest route and applied to the tourism of Banda Aceh city and Aceh Besar regency. In this reserach, Ant Colony Optimization algorithm is used to determine the shortest route to tourism of Banda Aceh city and Aceh Besar regency. From the analysis made by using both manual calculation and GUI MATLAB program application test, the shortest route can be obtained with a minimum distance of 120.85 km in one travel. Based on the test result, the application for tourism (in Banda Aceh city and Aceh Besar regency) shortest route searching built by utilizing the Ant Colony Optimization algorithm can find optimal route.