Flavin mononucleotide (FMN) derived from Vitamin B2, a bio-based fluorescent water-soluble molecule with visible yellow-green fluorescence, has been used in the scope of producing photoluminescent and glow-in-the-dark patterned polyester (PET) nonwoven panels. Since the FMN molecule cannot diffuse inside the PET fiber, screen printing, coating, and padding methods were used in an attempt to immobilize FMN molecules at the PET fiber surface of a nonwoven, using various biopolymers such as gelatin and sodium alginate as well as a water-based commercial polyacrylate. In parallel, air atmospheric plasma activation of PET nonwoven was carried for improved spreading and adhesion of FMN bearing biopolymer/polymer mixture. Effectively, the plasma treatment yielded a more hydrophilic PET nonwoven, reduction in wettability, and surface roughness of the plasma treated fiber with reduced water contact angle and increased capillary uptake were observed. The standard techniques of morphological properties were explored by a scanning electron microscope (SEM) and atomic force microscopy (AFM). Films combining each biopolymer and FMN were formed on PS (polystyrene) Petri-dishes. However, only the gelatin and polyacrylate allowed the yellow-green fluorescence of FMN molecule to be maintained on the film and PET fabric (seen under ultraviolet (UV) light). No yellow-green fluorescence of FMN was observed with sodium alginate. Thus, when the plasma-activated PET was coated with the gelatin mixture or polyacrylate bearing FMN, the intense photoluminescent yellow-green glowing polyester nonwoven panel was obtained in the presence of UV light (370 nm). Screen printing of FMN using a gelatin mixture was possible. The biopolymer exhibited appropriate viscosity and rheological behavior, thus creating a glow-in-the-dark pattern on the polyester nonwoven, with the possibility of one expression in daylight and another in darkness (in presence of UV light). A bio-based natural product such as FMN is potentially an interesting photoluminescent molecule with which textile surface pattern designers may create light-emitting textiles and interesting aesthetic expressions.