Laser shock peening (LSP) is a widely used surface treatment technique that can effectively improve the fatigue life and impact toughness of metal parts. Cr5Mo1V steel exhibits a gradient hardened layer after a LSP process. A new method is proposed to estimate the impact toughness that considers the changing mechanical properties in the gradient hardened layer. Assuming a linearly gradient distribution of impact toughness, the parameters controlling the impact toughness of the gradient hardened layer were given. The influences of laser power densities and the number of laser shots on the impact toughness were investigated. The impact toughness of the laser peened layer improves compared with an untreated specimen, and the impact toughness increases with the laser power densities and decreases with the number of laser shots. Through the fracture morphology analysis by a scanning electron microscope, we established that the Cr5Mo1V steel was fractured by the cleavage fracture mechanism combined with a few dimples. The increase in the impact toughness of the material after LSP is observed because of the decreased dimension and increased fraction of the cleavage fracture in the gradient hardened layer.