The congestion of pedestrians impedes the utilization efficiency of a subway station. Conflicts among pedestrians due to unseparated pedestrian flows not only increase the impedance of pedestrian mobility but also negatively affect on pedestrian safety. This paper analyzes the travel characteristics of bi-directional pedestrian flow based on microscopic movements, and evaluates the operation efficiency on separating the traffic line. The subway station was simulated in a 2-D grid structure by applying Discrete Element Method, and the movement is organized in each cell of the grid. As a result, the model explicates that separating the traffic line and encouraging the 'Keep right rule' would be mostly effective for the conflicting flows. Therefore, applying the 'Walking Guidance System' would be efficient to improve the pedestrian convenience and mobility.