ZnSe nanoparticles have been synthesized by microwave assisted method by using zinc chloride, selenium powder and ethylene diamine. The synthesized nanoparticles have been characterized structurally by FT-IR and XRD as well as morphological characterization was done by scanning electron microscope (SEM). The crystallite size after synthesis was obtained around 30 nm for pure ZnSe nanocrystallites. However, from SEM micrograph, agglomerated ZnSe nanoparticles of irregular shapes were observed. The assynthesized ZnSe nanoparticles at different contents (1 to 5% w/w w.r.t SPI) were incorporated into soy protein isolate (SPI) to produce reinforced SPI films by solution casting method. The ZnSe nanoparticles incorporated SPI suspensions were subjected to molecular mass and specific conductivity studies. Neat and ZnSe nanoparticles incorporated SPI films were structurally and mechanically characterized by FT-IR and tensile properties, respectively. Transmittance and water uptake studies were also carried out for ZnSe nanoparticles incorporated SPI films. The tensile strength and modulus increased from 5.80 MPa to 10.06 MPa and 18.84 MPa to 94.70 MPa with the increase in the contents of ZnSe nanoparticles from 0 to 5%. Moreover, the results also revealed a good antibacterial effect in ZnSe nanoparticles incorporated SPI film. The main application of nanoparticles incorporated SPI film will be in the area of biodegradable packaging.