Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) has attracted substantial attention as a promising material for industrial applications. In this study, different PHBV films with distinct 3-hydroxyvalerate (3HV) contents produced by Azotobacter vinelandii OP were evaluated. The 3HV fraction ranged from 18.6 to 36.7 mol%, and the number-average molecular weight (Mn) was between 238 and 434 kDa. In the bioreactor, a 3HV fraction (36.7 mol%) and an Mn value of 409 kDa were obtained with an oxygen transfer rate (OTR) of 12.5 mmol L− 1 h− 1 . Thermal analysis measurements showed decreased melting (Tm) and glass transition (Tg) temperatures, and values with relatively high 3HV fractions indicated improved thermomechanical properties. The incorporation of the 3HV fraction in the PHBV chain improved the thermal stability of the films, reduced the polymer Tm, and affected the tensile strength. PHBV film with 36.7 mol% 3HV showed an increase in its tensile strength (51.8 MPa) and a decrease in its Tm (170.61 ◦C) compared with PHB. Finally, scanning electron microscopy (SEM) results revealed that the PHBV film with 32.8 mol% 3HV showed a degradation upon contact with soil, water, or soil bacteria, showing more porous surfaces after degradation. The latter phenomenon indicated that thermomechanical properties played an important role in biodegradation.