As important tool for engine semi-automatic isolation system, the magneto-rheological fluids (MRC) can’t achieve tunable stiffness but damping. Therefore, the MRC’s initial stiffness affects the performance of isolation system. To obtain the better isolation effect of MRC in wide frequency, it is necessary to optimize the initial stiffness. As motor unbalanced force and isolation damping are not considered in traditional mount optimization, the feature of semi-automatic control can not be reflected. For a four-cylinder four-cycle in-line diesel engine, a new method which is suitable for stiffness optimization on vehicle engine isolation is presented by using Genetic Algorithm. The optimal isolation system has small absolute force transmissibility in abroad frequency range and small vibration intensity with impact load by using the new method.