Recently, due to sustainable development and environmental protection policies, there is increasing interest in the development of new biodegradable polymer-based multifunctional composites. Chitosan is one of the most remarkable and preferred biopolymers, which is environmentally friendly as well as renewable, biocompatible, and inexpensive. Though it has a wide range of potential applications, the major limitation of chitosan – the problem of poor mechanical performance – needs to be solved. In this work, graphene oxide was first produced and then used to manufacture a chitosan/graphene oxide/zinc oxide composite film through a casting method. The properties of the chitosan film and the chitosan/graphene oxide/zinc oxide composite film were investigated using Fourier transform infrared spectroscopy, mechanical, thermal gravimetric, and ultraviolet (UV)-visible spectroscopy analyses. The results showed that the incorporation of graphene oxide and zinc oxide into the chitosan matrix resulted in enhanced mechanical properties and thermal stability of chitosan biocomposite films. The graphene oxide- and zinc oxide-reinforced chitosan film showed 2527 MPa and 55.72 MPa of Young’s modulus and tensile strength, respectively, while neat chitosan showed only 1549 MPa and 37.91 MPa of Young’s modulus and tensile strength, respectively. Conversely, the addition of graphene oxide decreased the transmittance, notably in the UV region.