Assembly U-lines are increasingly accepted in industry, especially just-in-time production systems, for the efficient utilization of workforce. In this paper, we present an integer programming formulation and a branch-and-bound method for balancing the U-line with the objective of minimizing the number of workstations with a fixed cycle time. In the mathematical model, we provide the method that can reduce the number of variables and constraints. The proposed branch-and-bound method searches the optimal solution based on a depth-first-search. To efficiently search for the optimal solutions to the problems, an assignment rule is used in the method. Bounding strategies and dominance rules are also utilized. Some problems require a large amount of computation time to find the optimal solutions. For this reason. some heuristic fathoming rules are also proposed. Extensive experiments with test-bed problems in the literature are carried out to show the performance of the proposed method. The computational results show that our method is promising in solution quality.