Starch (PS) isolated from fresh pasta by‐product was plasticized with different amount of glycerol (10, 15, and 20%). X‐ray diffraction highlighted that after PS plasticization, the obtained bio‐films showed a partial change in the crystallinity and typical saccharide bands were put in evidence by peaks associated with C‐C, C‐O, C‐H, and ‐OH stretching from the Fourier transform infrared spectroscopy spectra. The differential scanning calorimetry measurements demonstrated that glass transition temperature (Tg) of bio‐films decreased with increasing glycerol content. In addition, their total soluble matter released in distilled water increased with increasing the glycerol content; and an increase in relative humidity and in plasticizer content resulted in the highest values of water uptake. The tensile properties revealed a significant decrease in both Young's modulus and tensile strength but an increase in elongation at break with increasing the glycerol content. The biodegradation tests proved that bio‐films were more biodegradable than the extracted starch implying a strong connection between plasticizer and biodegradability. © 2018 American Institute of Chemical Engineers Environ Prog, 38: 527–533, 2019