Bus passenger transport is very important for sustainable urban mobility, and due to the characteristics of the street network, it is usually realized in the conditions of mixed traffic flow. Disturbances and sudden changes of traffic flow parameters occurring in the street network in a mixed traffic flow affect the disruptions in schedule and travel time of all vehicles in the traffic flow, including public transport (PT) vehicles. In order to keep to the planned schedule in the peak hours of PT lines carried out in the conditions of mixed traffic flow, the operators introduce new vehicles or reduce the vehicles’ dwell time at terminuses, which is often impossible to do. The use of a larger number of public transport vehicles increases the fuel consumption, pollutants’ emission, and the operating costs. In this paper, a network optimization model was developed for defining the influence of the change of traffic flow parameters in a mixed traffic flow on travel time of PT vehicles. The model takes into consideration uncertain time unevenness of the change of traffic flow parameters, which enables determining the optimization of travel time and defining the necessary number of public transport vehicles for the purpose of keeping to the planned schedule. In order to develop the transport model, counting and analysis of the characteristics of traffic flow at 61 intersections on the city territory were carried out. The model was tested on bus line number 4 of PT in Novi Sad. The model showed that it is possible to achieve certain savings regarding the number of vehicles with the unchanged headway, that is, the unchanged level of service which is offered to the users. With the application of the model in real traffic conditions, significant savings, as well as operating and external costs’ reduction, can be achieved, which contributes to the sustainability of public bus transport in urban environments.