Bio-catalysts are non-toxic, metal-free, abundant, and efficient materials with less waste generation that decrease the environmental pollution. Nano-ovalbumin is a biodegradable phosphoglyco protein with isoelectric point (pI) of 4.5, molecular weight of 44.5 kDa containing 385 residues of amino acids. Nano-ovalbumin can be used as a non-toxic heterogeneous and eco-friendly catalyst for promotion of organic compounds. Quinazolinones have many biological and medicinal activities such as anticonvulsant, antitumor, anticancer, hypnotic/sedatives, antimalarial, antibacterial, antioxidant, antimicrobial, antifungal, cytotoxic, antiproliferative, and inhibitory activities on α-glucosidase. In this research, nano-ovalbumin was ready by a new, easy, inexpensive and convenient protocol from egg white and used as a heterogeneous biocatalyst in the synthesis of 2,3-dihydroquinazolin-4(1H)-ones via two-component reaction of 2-aminobenzamide with aldehyde or ketone. This eco-friendly catalyst carried out the reaction in green conditions and with good to excellent yields. The structure of products were determined by Fourier Transform-Infrared (FT-IR), 1 H-Nuclear Magnetic Resonance ( 1 H-NMR) and their physical properties were compared with those reported in others work.