In this study, we designed and fabricated a poling-free piezoelectric nanogenerator using bio-based polymer and filler, a poly(l-lactic acid) (PLLA)/vitamin B-2(VB2) composite film. The addition of VB(2)to PLLA led to improved crystallinity and enhanced the molecular orientation of PLLA, resulting in a significantly enhanced output performance of the PLLA/VB(2)nanogenerator. The proposed nanogenerator provides a maximum power density of 75.68 mW cm(-3), which is approximately 28.7 times higher than that of pure PLLA nanogenerator. Moreover, the PLLA/VB(2)nanogenerator exhibits good electrical responses under both mechanical and acoustic stimulations. The collected mechanical energy from the motion of human joints can power up several LEDs. The PLLA/VB(2)nanogenerator can sensitively detect sound frequency from 1 to 27 kHz. Thus, it has great application potential in ambient mechanical and acoustic energy harvesting, wearable devices, and human-machine controlling systems.