Intelligent manufacturing workshop uses automatic guided vehicles as an important logistics and transportation carrier, and most of the existing research adopts the intelligent manufacturing workshop layout and Automated Guided Vehicle (AGV) path step-by-step optimization, which leads to problems such as low AGV operation efficiency and inability to achieve the optimal layout. For this reason, a smart manufacturing assembly line layout optimization model considering AGV path planning with the objective of minimizing the amount of material flow and the shortest AGV path is designed for the machining shop of a discrete manufacturing enterprise of a smart manufacturing company. Firstly, the information of the current node, the next node and the target node is added to the heuristic information, and the dynamic adjustment factor is added to make the heuristic information guiding in the early stage and the pheromone guiding in the later stage of iteration; secondly, the Laplace distribution is introduced to regulate the volatilization of the pheromone in the pheromone updating of the ant colony algorithm, which speeds up the speed of convergence; the path obtained by the ant colony algorithm is subjected to the deletion of the bi-directional redundant nodes, which enhances the path smoothing degree; and finally, the improved ant colony algorithm is fused with the improved dynamic window algorithm, so as to enable the robots to arrive at the end point safely. Simulation shows that in the same map environment, the ant colony algorithm compared with the basic ant colony algorithm reduces the path length by 40% to 67% compared to the basic ant colony algorithm and reduces the path inflection points by 34% to 60%, which is more suitable for complex environments. It also verifies the feasibility and superiority of the conflict-free path optimization strategy in solving the production scheduling problem of the flexible machining operation shop.