This study aimed to incorporate pistachio shell hemicellulose into a film of gelatin and glycerol for the production of biodegradable films. The gelatin and glycerol are chosen because of their functional properties, which make it extensively used in food industry. The film composition was defined after a statistical optimization by central composite face-centered design and response surface methodology. The hemicellulose/gelatin ratio of 35.93% and the glycerol ratio of 18.02% were the optimum conditions to obtain lower film water solubility, higher tensile strength, and elongation at break values. The physical, structural, mechanical, and barrier properties of the developed hemicellulose-gelatin film were analyzed and compared with those of the gelatin film. Tensile strength and film water solubility values were reduced significantly with hemicellulose incorporation from 20.41 to 16.64 MPa and 49.57 to 39.21%, respectively, while EB was enhanced by 4.34 times. In addition, hemicellulose incorporation enhanced the water vapor permeability and the film degradation in the soil. The films were also examined by Fourier transform infrared spectroscopy and differential scanning calorimetry. The novelty of this study is to use pistcahio shell hemicellulose in the production of an edible film for the first time.