Nowadays, the optimization methods are widely used to adjust controller parameters to tune their optimal values in order to enhance the efficiency and performance of dynamic systems. In this study, the parameters of a linear PI controller were optimized by using five different optimization algorithms such as Artificial Tree Algorithm (ATA), Particle Swarm Optimization (PSO), Differential Evolution Algorithm (DEA), Constrained Multi-Objective State Transition Algorithm (CMOSTA), and Adaptive Fire Forest Optimization (AFFO). The optimized controllers were implemented in real time for temperature control of a Heat-Flow System (HFS) under various step and time-varying reference signals. In addition, the Ziegler–Nichols (Z-N) method was also applied to the system as a benchmark to compare the temperature tracking performance of the proposed optimization methods. To further evaluate the performance of each optimization algorithm, Mean Absolute Error (MAE) values were calculated and improvement ratios were obtained. The experimental results showed that the proposed optimization methods provided more successful reference tracking and enhanced controller performance as well.