Manual harvesting of white button mushrooms involves coordinated bending and twisting motions to detach the fruiting body while minimizing surface damage; however, replicating these actions in automated systems remains challenging. In this study, a vacuum-based end-effector that mimics manual twist–bend detachment using a single-point contact was designed and evaluated to reduce mechanical damage. Key detachment parameters, including the friction coefficient (mean 0.62), bending angle (average 5.72°), and twisting torque (average 2.56 N·m), were experimentally analyzed to determine the minimum vacuum pressures required for effective bending and twisting, which were −8.64 ± 2.21 kPa and −8.91 ± 2.45 kPa, respectively, with no significant difference observed between the two motions (p = 0.51). A customized vision-based image processing algorithm was developed to quantify postharvest surface damage using a whiteness index (WI). An optimal vacuum pressure of −17.17 kPa was identified, together with a bending angle of 10° and a twisting angle of 90°, balancing high harvesting success with preservation of mushroom quality. The results highlight the influence of end-effector design parameters, including vacuum cup material, contact area, bending direction, and vacuum application duration, on harvesting performance and product marketability, supporting the development of robotic systems for fresh mushroom harvesting.