In this study, a piezoelectric nanogenerator (PENG) based on multilayer composite fiber had good and stable piezoelectric output performance, which could realize biomechanical energy collection and human movement monitoring. The polyvinylidene fluoride‐hexafluoryl propylene (P(VDF‐HFP)) composite fiber doped with MXene was used as a promising piezoelectric functional layer (MPFP). By electrospinning, P(VDF‐HFP) composite fiber was constructed by alternating electrospinning with polyurethane (TPU) nanofibers layer by layer. The adoption of MXene as a functional filler promoted the transformation of P(VDF‐HFP) from α to piezoelectric β‐crystal, the piezoelectric β‐crystal content of P(VDF‐HFP) increased, and the dielectric properties and polarization levels were enhanced. At the same time, the multi‐layer structure design improved the piezoelectric sensitivity and mechanical properties of the composite fiber, and the composite fiber was more flexible and had longer tensile strain. With excellent dielectric and mechanical properties, 3MPFP/TPU/3MPFP/TPU multilayer composite fibers showed piezoelectric sensitivity of 10.88 V/kPa. Under the action of palm pressing, the PENG generated an open circuit voltage of 25 V, and the voltage signal of the device under different motion forms had specific characteristics such as peak value, frequency, and shape, which could be used to realize the analysis of human motion forms. This study provided an efficient, simple, and creative new idea for the application of flexible energy storage electronic devices, PENGs, and piezoelectric sensors.