Bioluminescent organisms represent a largely untapped resource at the interface of biodiversity, bioentrepreneurship, and sustainable ecotourism. Although bioluminescence is well studied biologically, its translational potential for commercial, educational, and tourism-based applications has not been systematically synthesized. This review addresses this gap by evaluating the opportunities, limitations, and ethical considerations associated with the use of wild, laboratory-cultured, and transgenic bioluminescent organisms. Using published literature, case studies, and applied examples to assess current and emerging applications in bioentrepreneurship (aquarium and gift markets, biosensors, transgenic lighting, art, education, film, and culinary novelty products) and ecotourism (natural bioluminescent hotspots, laboratory-based exhibits, and deep sea viewing). This review identifies key opportunity domains, including scalable planktonic and microbial systems for year-round use, laboratory-controlled tourism and education platforms, and high-value niche ecotourism such as firefly festivals and glowing beaches. Major challenges include limited baseline ecological data, difficulties in culturing and scaling luminous organisms, seasonal and spatial unpredictability, regulatory and ethical constraints, infrastructure limitations, and risks of habitat disturbance, particularly for rare or endemic organisms. This review infers that bioluminescence-based entrepreneurship and ecotourism can support sustainable bioeconomies, conservation awareness, and local livelihoods when guided by robust science, ethical governance, community participation, and long-term ecological monitoring.