This paper presents the design and implementation of a novel control scheme for frequency regulation in an AC microgrid, subjected to varying operating conditions, using fractional order proportional integral derivative (FOPID) controller with its parameters optimized employing multi-verse optimizer (MVO) evolutionary algorithm. For the comparative analysis, proportional integral derivative (PID) and fractional order proportional integral (FOPI) controllers, both optimally tuned using MVO, are also implemented. Further, the impact of electric vehicle (EV) on system performance is analyzed. Integral of time multiplied absolute error (ITAE) is considered as the cost function in the optimization process whereas, the settling time and peak undershoot serve as the performance indices and a step change in load is taken as the perturbation. The proposed control scheme is tested for robustness against system parametric changes in the range of ± 25 besides random step load varying both in magnitude and time scale. The proposed MVO optimized FOPID controller outperforms other control schemes implemented in this work and proves to be very effective in frequency regulation in the AC microgrid comprising the renewable energy sources (RESs), diesel engine generator (DEG), and the energy storage systems (ESS). System Modeling and simulations are performed in MATLAB/Simulink.